Plants and seeds of brassica carinata variety dh-040.008

ABSTRACT

The invention is in the field of  Brassica carinata  breeding (i.e. Ethiopian mustard breeding), specifically relating to  Brassica carinata  variety DH-040.008. The present invention relates to seeds, plants or parts thereof, cells, methods of making, and uses of this variety and its progeny.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of and priority from U.S.Provisional Patent Application No. 62/689,402 filed Jun. 25, 2018, thecontents of which are incorporated herein by reference in theirentirety.

FIELD OF THE INVENTION

The present invention relates generally to the field of Brassicacarinata breeding and, more specifically, to the development of a newBrassica carinata variety DH-040.008, a sample of seed of which has beendeposited with NCIMB under Accession number 42999.

BACKGROUND

Brassica carinata is a member of the Brassicaceae (formerly Cruciferae)family, commonly known as the mustard family. In Canada, Brassicacarinata is commonly known as carinata, but is also sometimes referredto as Ethiopian mustard, Abyssinian mustard, Ethiopian kale orAbyssinian kale. In Ethiopia it is named gomenzer (Getinet, et al.,1996).

The genus Brassica is a member of the tribe Brassiceae in the mustardfamily (Brassicaceae; (Warwick, et al., 2009). In addition to B.carinata, the Brassica genus includes several economically importantoilseed crop species: B. juncea (L). Czern. (brown mustard), B. napus L.(rape, Argentine canola), B. nigra (L.) W. D. J. Koch (black mustard),and B. rapa L. (field mustard, Polish canola). The genus Brassica alsoincludes B. oleracea L. food crops, including cabbage, broccoli,cauliflower, brussels sprouts, kohlrabi and kale.

The six Brassica species are closely related genetically, as describedin the Triangle of U (Nagaharu, 1935). Brassica carinata is anamphidiploid (BBCC, 2n=34) thought to be derived from interspecifichybridization of the diploid species B. nigra L. (BB, 2n=16) and B.oleracea L. (CC, 2n=18; (Prakash, et al., 2011). The native range ofBrassica carinata comprises the central highland region of Ethiopia. Allthe naturally occurring carinata in these regions is cultivated; theredo not appear to be wild populations.

Brassica carinata is an herbaceous annual with a determinate growthhabit (Zanetti, et al., 2013). The plants were originally cultivated intheir home range primarily as a source of leaves used as an edible andnutritious vegetable (Neugart, et al., 2017). Carinata can be grown as acover crop to reduce soil erosion and herbicide use, while promotingwater conservation (Alcantara, et al., 2011) or can be plowed into thesoil for use as a green manure soil amendment and bio-fumigant (Lazzeri,et al., 2009; Pane, et al., 2013). Carinata also has utility inphytoremediation of heavy metal-contaminated topsoil (Mourato, et al.,2015). As abundant producers of vegetative biomass, cropping ofcarinata's above-ground biomass has been suggested as a renewable sourceof feedstock for conversion to energy, particularly where cultivated insouthern Europe (Gasol, et al., 2007).

Brassica carinata can be grown in subtropical regions as winter covercrop in rotations with summer crops such as beans, cotton, and peanuts,where the usual practice had been to follow with winter fallow. This ismade possible by the unique ability for established carinata to surviveand recover after hard frosts (Seepaul, et al., 2015). Benefits ofcarinata's use as a winter cover crop in this environment include theability to conserve winter moisture and nutrients in the soil, mitigateleaching of nitrogen, phosphates and other residual nutrients into localwaterways, as well as providing a means to increase soil organic carbon(Newman, et al., 2010 (revised)). Brassica carinata's greater toleranceto early season frost, ability to better cope with higher heat and lowermoisture during flowering and seed set, as well as resistance tolodging, allows it to better withstand early and late season weatherextremes (Seepaul, et al., 2015), making it overall a more reliableoilseed cropping option for producers in semi-arid regions.

Brassica crops have long been shown to be beneficial when grown inrotations with cereals such as wheat, an important food crop amenable toproduction in semiarid regions by virtue of its shorter growing seasonand tolerance to climate extremes. Rotations with oilseed as well asforage Brassica species have consistently demonstrated a beneficialeffect on yield of the ensuing cereal crop, due to their effects onimproving soil structure and moisture conservation and to its ability toprovide a break to the cycle of diseases that affect cereal performance(Angus, et al., 2011). Its ability to break cereal disease cycles stemsfrom Brassica's lack of susceptibility to many cereal diseases, but mayalso derive from their ability to actively discourage persistence ofsoil pathogens via the biofumigant activity of root exudates andresidues (Kirkegaard and Sarwar, 1998). In the southern hemisphere, thecrop can be sown in late autumn or early winter into moist soil. Inhigher rainfall zones, it can be sown as late as early spring.

In terms of economic value, Brassica carinata's greatest potential as acrop resides in its prolific yields of oil and protein rich seed. Insouthern Europe, carinata seed oil has been investigated for itspotential as a feedstock for biofuel and as a bio-industrial feedstockwith applications in production of lubricants, paints, cosmetics,plastics (Cardone, et al., 2002; Cardone, et al., 2003; Bouaid, et al.,2005; Gasol, et al., 2007; Gasol, et al., 2009). In North America, wherevarieties have been adapted to grow in regions as diverse as thesemi-arid southern Canadian prairies and adjacent US northern tierstates as well as the Southeast US gulf states, carinata has been shownto be a suitable renewable feedstock crop for biofuel production (Gesch,et al., 2015; Seepaul, et al., 2015), and oil extracted from B. carinataseed has been used to produce green bio-diesel and bio-jet fuel (Drenth,et al., 2015). In October 2012, experimental aviation flights by theNational Research Council of Canada using the world's first 100% bio-jetfuel were successful (“ReadiJet 100% biofuels flight—one of 2012's 25most important scientific events”, Popular Science Magazine, 2012(12).

Carinata varieties have been developed that are optimized for productionof oil feedstock for diverse bio-industrial uses such as manufacturingof bio-plastics (Impallomeni, et al., 2011; Newson, et al., 2014),lubricants (Zanetti, et al., 2009) and specialty fatty acids such as 5,13-docosadienoic acid, 5-eicosenoic acid (Jadhav, et al., 2005),eicosapentaenoic acid (Cheng, et al., 2010) and nervonic acid (Taylor,et al., 2010). In some cases, modification of the seed oil profile hasinvolved the use of transgenic technologies to introduce specific genesencoding enzymes of the fatty acid biosynthesis pathways or constructsto knock down expression of endogenous pathway genes (reviewed inTaylor, et al., 2010).

As well as its high oil content, carinata seed has high protein and lowfibre content (Xin and Yu, 2014), making the meal that is produced as aby-product of the oil extraction process a potential source of highquality protein for use in animal feed applications. The native seedalso has a high content of glucosinolate (GSL), a class of sulfurcontaining compounds which, when present at high levels in meal, canreduce feed palatability and adversely affect animal health. There havebeen efforts to reduce GSL levels through use of interspecific crossingwith low GSL varieties of other Brassicaceae species (Getinet, et al.,1997; Márquez-Lema, et al., 2008). More recently, it has beendemonstrated that the optimized processing of the meal during the oilextraction process can remove most of the glucosinolate, rendering themeal suitable for a number of animal feed applications (U.S. PublicationNo. 2018/004226). Carinata meal is currently approved as a supplementfor cattle feed in Canada, US and Europe.

Brassica carinata has been adapted to meet the demands of emergingmarkets for carinata seed and carinata products. To provide opportunityto expand its production base, there continues to be a great need in theart for new carinata varieties and lines with improved traits, includingincreased grain and oil yields per acre, increased seed oil content andoptimized oil composition, as well as varieties with improved agronomictraits such as increased tolerances to biotic and abiotic stresses,reduced time to maturity and improved harvestability.

SUMMARY OF THE INVENTION

In one aspect of the invention, a new Brassica carinata varietyDH-040.008 is provided. In other aspects, the invention also provides aseed, plant, plant part or cell of Brassica carinata variety DH-040.008,for which a representative sample of seed has been deposited under NCIMBAccession Number 42999.

In other aspects, the invention provides methods for producing aBrassica carinata plant by crossing variety DH-040.008 with itself orwith other Brassica carinata varieties. The invention is also directedto a cell, seed, plant, or plant part of a Brassica carinata varietyproduced by crossing Brassica carinata variety DH-040.008 with itself orwith other Brassica carinata varieties.

In another aspect, the invention is directed to a Brassica carinata seedproduced by crossing Brassica carinata plants and harvesting theresulting Brassica carinata seed, wherein at least one Brassica carinataplant is the plant of Brassica carinata variety DH-040.008. In anotheraspect, the invention is directed to a method of producing a Brassicacarinata variety derived from Brassica carinata variety DH-040.008, themethod comprising (a) crossing Brassica carinata variety DH-040.008plant with a different Brassica carinata plant having a desired trait toproduce F1 hybrid seed; and (b) growing the resultant F1 hybrid seed andselecting one or more F1 hybrid progeny plants having the desired traitand at least a portion of the genetic make up of Brassica carinatavariety DH-040.008. In one embodiment, the method of producing aBrassica carinata variety derived from Brassica carinata varietyDH-040.008 further comprises the steps of (a) backcrossing the selectedprogeny plants with plants of variety DH-040.008, or with the differentBrassica carinata plant having a desired trait, to produce backcrossprogeny seed; (b) growing the resultant backcross progeny seed andselecting backcross progeny plants that have the desired trait and atleast a portion of the genetic make up of Brassica carinata varietyDH-040.008; and (c) repeating steps (a) and (b) on the selectedbackcross progeny plants to a maximum of 10 generations to produce aprogeny Brassica carinata plant derived from Brassica carinata varietyDH-040.008, wherein the progeny Brassica carinata plant comprises thedesired trait and at least a portion of the genetic make up of Brassicacarinata variety DH-040.008. In another embodiment, the method ofproducing a Brassica carinata variety derived from Brassica carinatavariety DH-040.008 further comprises the steps of (a) self-pollinatingthe selected F1 hybrid progeny plants to produce further progeny seed;(b) growing the further progeny seed and selecting further progenyplants that have the desired trait and at least a portion of the geneticmake up of Brassica carinata variety DH-040.008; and (c) repeating steps(a) and (b) on the selected further progeny plants to a maximum of 10generations to produce a progeny Brassica carinata plant derived fromBrassica carinata variety DH-040.008, wherein the progeny Brassicacarinata plant comprises the desired trait and at least a portion of thegenetic make up of Brassica carinata variety DH-040.008. The inventionis also directed to a cell, seed, plant, or plant part of a Brassicacarinata variety derived from Brassica carinata variety DH-040.008 usingthe above-described methods.

In another aspect, the invention provides methods for producing doubledhaploid (DH) varieties from F1 Brassica carinata plants produced bycrossing Brassica carinata DH-040.008 variety with itself or with otherBrassica carinata varieties, as well as a cell, seed, plant, or plantpart produced by such DH varieties and any progeny of these.

In another aspect, the invention provides methods for producing aBrassica carinata plant by outcrossing Brassica carinata varietyDH-040.008 with other Brassicaceae species followed by backcrossing withBrassica carinata variety DH-040.008, as well as producing DH varietiesfrom the interspecific crosses. In some embodiments, the otherBrassicaceae species may be any species of the family Brassicaceaeincluding but not limited to Brassica alba, Brassica hirta, Brassicajuncea, Brassica napus, Brassica nigra, Brassica oleracea, Brassicarapa, Sinapus alba, and Camelina sativa. In some embodiments, theprogeny DH varieties retain the “essential morphological orphysiological characteristics” of Brassica carinata variety DH-040.008as described herein, when grown in the same location under the sameenvironmental conditions. In some embodiments, the “essentialmorphological or physiological characteristics” of Brassica carinatavariety DH-040.008 are the physiological and/or morphologicalcharacteristics set forth in one or more of Tables 1, 2, 3, 4, and 5 asdetermined at the 5% significance level.

In another aspect, the invention provides for the use of Brassicacarinata variety DH-040.008 as a background for chemical and/orradiation induced mutagenesis, for targeted gene editing, for modulationof traits via RNA interference or antisense RNA expression, or forintroduction of traits via genetic transformation.

In another aspect, the invention provides a method to produce acommercial crop of plants of Brassica carinata variety DH-040.008, aswell any progeny plants derived from Brassica carinata varietyDH-040.008. The invention also provides a method of producing acommercial plant product comprising growing the plant of Brassicacarinata variety DH-040.008, or any progeny plants derived from Brassicacarinata variety DH-040.008, to produce a commercial crop and producingthe commercial plant product from the commercial crop.

In other aspects, the invention relates to the use of Brassica carinatavariety DH-040.008 to produce a commercial product, such as oil, meal,protein isolate, biofumigant, or crushed non-viable seed. The inventionalso provides for commercial products produced from Brassica carinatavariety DH-040.008, as well any progeny plants derived from Brassicacarinata variety DH-040.008, including but not limited to oil, meal,protein isolate, biofumigant, or crushed non-viable seed.

The invention provides for, without limitation, the following numberedembodiments:

-   1. Brassica carinata variety DH-040.008, representative seed of the    variety having been deposited under NCIMB accession number 42999.-   2. A seed, plant, plant part or cell of Brassica carinata variety    DH-040.008, representative seed of the variety having been deposited    under NCIMB accession number 42999.-   3. A Brassica carinata seed produced by crossing Brassica carinata    plants and harvesting the resulting Brassica carinata seed, wherein    at least one Brassica carinata plant is the plant of embodiment 2.-   4. A method of producing a Brassica carinata variety derived from    Brassica carinata variety DH-040.008, the method comprising    -   (a) crossing a plant of Brassica carinata variety DH-040.008        with a different Brassica carinata plant having a desired trait        to produce F1 hybrid seed; and    -   (b) growing the resultant F1 hybrid seed and selecting one or        more F1 hybrid plants having the desired trait and at least a        portion of the genetic make up of Brassica carinata variety        DH-040.008.-   5. The method of embodiment 4, further comprising the steps of    -   (a) backcrossing the selected F1 hybrid plants with plants of        Brassica carinata variety DH-040.008, or with the different        Brassica carinata plant having a desired trait, to produce        backcross progeny seed;    -   (b) growing the resultant backcross progeny seed and selecting        backcross progeny plants that have the desired trait and at        least a portion of the genetic makeup of Brassica carinata        variety DH-040.008; and    -   (c) repeating steps (a) and (b) on the selected backcross        progeny plants to a maximum of 10 generations to produce a        Brassica carinata plant derived from Brassica carinata variety        DH-040.008, wherein the resulting progeny Brassica carinata        plant comprises the desired trait and at least a portion of the        genetic makeup of Brassica carinata variety DH-040.008.-   6. The method of embodiment 4, further comprising the steps of    -   (a) self-pollinating the F1 hybrid plants to produce further        progeny seed;    -   (b) growing the further progeny seed and selecting further        progeny plants that have the desired trait and at least a        portion of the genetic make up of Brassica carinata variety        DH-040.008; and    -   (c) repeating steps (a) and (b) on the selected further progeny        plants to a maximum of 10 generations to produce a Brassica        carinata plant derived from Brassica carinata variety        DH-040.008, wherein the Brassica carinata plant comprises the        desired trait and at least a portion of the genetic makeup of        Brassica carinata variety DH-040.008.-   7. A cell, seed, plant, or plant part of the Brassica carinata    variety produced by the method of any one of embodiments 4 to 6,    wherein the Brassica carinata plant has the same physiological    and/or morphological characteristics set forth in one or more of    Tables 1, 2, 3, 4, and 5, as determined at the 5% significance    level, when grown in the same location under the same environmental    conditions as variety DH-040.008 as a plant of variety DH-040.008.-   8. A method of producing a commercial plant product, the method    comprising growing the plant of embodiment 2 to produce a commercial    crop and producing the commercial plant product from the commercial    crop.-   9. The method of embodiment 8, wherein the commercial plant product    comprises oil, meal, protein isolate, or biofumigant.-   10. A method of producing a commercial plant product, the method    comprising growing the plant of embodiment 7 to produce a commercial    crop and producing the commercial plant product from the commercial    crop.-   11. The method of embodiment 10, wherein the commercial plant    product comprises oil, meal, protein isolate, or biofumigant.-   12. A commercial crop produced from the Brassica carinata plant of    embodiments 2 or 7.-   13. A commercial plant product produced from the Brassica carinata    plant of embodiments 2 or 7.-   14. The commercial plant product of embodiment 13, wherein the    commercial plant product comprises oil, meal, protein isolate or    biofumigant.-   15. The plant part of embodiment 2, wherein the plant part is an    ovule, a leaf, pollen, a seed, an embryo, a root, a root tip, a pod,    a flower, a stalk, a cell, or a protoplast.-   16. The plant part of embodiment 15, wherein the plant part is    pollen.-   17. The plant part of embodiment 15, wherein the plant part is an    ovule.-   18. A Brassica carinata plant or plant part having essentially all    the physiological and morphological characteristics of the plant of    embodiment 2 when grown in the same location under the same    environmental conditions.-   19. A Brassica carinata plant or plant part having the physiological    and/or morphological characteristics set forth in one or more of    Tables 1, 2, 3, 4, and 5, as determined at the 5% significance    level, when grown in the same location under the same environmental    conditions as variety DH-040.008.-   20. A method for producing Brassica carinata seed comprising    crossing Brassica carinata plants and harvesting the resulting    Brassica carinata seed, wherein at least one Brassica carinata plant    is the plant of embodiment 2.-   21. A method for producing a first generation (F1) hybrid Brassica    carinata seed comprising crossing the plant of embodiment 2 with a    different Brassica carinata plant and harvesting the resultant F1    hybrid Brassica carinata seed, and wherein the plant of embodiment 2    is either a female parent or a male parent.-   22. An F1 hybrid seed produced by the method of embodiment 21.-   23. An F1 hybrid plant grown from the F1 hybrid seed of embodiment    22.-   24. A method for producing a Double Haploid variety comprising:    -   (a) isolating a flower bud of the F1 plant of embodiment 23;    -   (b) dissecting out a haploid microspore;    -   (c) placing the haploid microspore in culture;    -   (d) inducing the microspore to differentiate into an embryo and        subsequently into a plantlet;    -   (e) identifying whether the plantlet contains a diploid        chromosome number, wherein the diploid chromosome number        occurred through chromosome doubling; and    -   (f) continuing to grow the plantlet if it contains a diploid        chromosome number, wherein the Double Haploid variety comprises        at least a portion of the genetic makeup of Brassica carinata        variety DH-040.008.-   25. The method of embodiment 24 further comprising inducing    chromosome doubling by chemical or physical means.-   26. A cell, plant, plant part, or seed of a Doubled Haploid variety    produced by the method of embodiment 24 or 25.-   27. A method of producing a Brassica carinata variety derived from    the plant of embodiment 2, wherein the Brassica carinata variety    comprises a desired trait, the method comprising the steps of:    -   (a) crossing a plant of variety DH-040.008 with another Brassica        carinata variety comprising the desired trait;    -   (b) growing the resultant F1 hybrid seed and selecting one or        more progeny plants that have the desired trait;    -   (c) backcrossing the selected progeny plants that have the        desired trait with plants of variety DH-040.008 to produce        backcross progeny seed; and    -   (d) growing the resultant backcross progeny seed and selecting        backcross progeny plants that have the desired trait,    -   wherein the Brassica carinata variety derived from the plant of        embodiment 2 comprises at least a portion of the genetic makeup        of Brassica carinata variety DH-040.008-   28. The method of embodiment 27, wherein steps (c) and (d) are    repeated until the Brassica carinata variety produced from variety    DH-040.008 has the desired trait and the physiological and/or    morphological characteristics set forth in one or more of Tables 1,    2, 3, 4, and 5, as determined at the 5% significance level, when    grown in the same location under the same environmental conditions    as variety DH-040.008.-   29. A method of producing a Brassica carinata variety from the plant    of embodiment 2, wherein the Brassica carinata variety comprises a    desired trait, the method comprising introducing a nucleic acid    construct conferring the desired trait into a Brassica carinata    plant of variety DH-040.008.-   30. The method of embodiment 29, wherein the nucleic acid construct    is introduced using polyethylene glycol (PEG) mediated nucleic acid    uptake, electroporation, ballistic infiltration using nucleic acid    coated microprojectiles (gene gun), an Agrobacterium    infiltration-based vector, or a plant virus based vector.-   31. The method of embodiment 29 or 30, wherein the nucleic acid    construct comprises a transgene, an RNAi construct, or an antisense    RNA construct.-   32. The method of any one of embodiments 29 to 31, wherein the    Brassica carinata variety comprises the desired trait and the    physiological and/or morphological characteristics set forth in one    or more of Tables 1, 2, 3, 4, and 5, as determined at the 5%    significance level, when grown in the same location under the same    environmental conditions as variety DH-040.008.-   33. A method of producing a Brassica carinata variety from the plant    of embodiment 2, wherein the Brassica carinata variety comprises a    desired trait, the method comprising the steps of:    -   (a) crossing a plant of variety DH-040.008 with another Brassica        carinata variety comprising the desired trait;    -   (b) growing the resultant F1 hybrid seed and selecting one or        more progeny plants that have the desired trait;    -   (c) self-pollinating the progeny plants that have the desired        trait to produce further progeny seed; and    -   (d) growing the further progeny seed and selecting further        progeny plants that have the desired trait,    -   wherein the Brassica carinata variety derived from the plant of        embodiment 2 comprises at least a portion of the genetic makeup        of Brassica carinata variety DH-040.008.-   34. The method of embodiment 33, wherein steps (c) and (d) are    repeated until the Brassica carinata variety from variety DH-040.008    has the desired trait the physiological and/or morphological    characteristics set forth in one or more of Tables 1, 2, 3, 4, and    5, as determined at the 5% significance level, when grown in the    same location under the same environmental conditions as variety    DH-040.008.-   35. A method of producing a Brassica carinata variety produced from    the plant of embodiment 2, wherein the Brassica carinata variety    comprises a new trait and the method comprises exposing seedlings or    microspores to a mutagenic agent and allowing the surviving fraction    to develop into mature plants.-   36. The method of embodiment 35, wherein the mutagenic agent is    ethyl methanesulfonate, N-ethyl-N-nitrosourea, or x-ray, gamma or    ultraviolet radiation.-   37. A method of producing a Brassica carinata variety derived from    the plant of embodiment 2, wherein the Brassica carinata variety    comprises a desired trait, the method comprising:    -   (a) crossing a plant of variety DH-040.008 with a plant of        another species of the family Brassicaceae comprising the        desired trait;    -   (b) producing F1 plants using embryo rescue techniques to        recover viable F1 plants or growing F1 seeds;    -   (c) self-pollinating the F1 plants that have the desired trait        and carinata character;    -   (d) producing F2 plants using embryo rescue techniques to        recover viable F2 plants or growing F2 seeds;    -   (e) self-pollinating the F2 plants that have the desired trait        and carinata character;    -   (f) producing F3 plants using embryo rescue techniques to        recover viable F2 plants or growing F3 seeds;    -   (g) self-pollinating the progeny plants that have the desired        trait and carinata character to produce further progeny plants;        and    -   (h) selecting further progeny plants with the desired trait and        carinata character to produce the carinata variety,    -   wherein the Brassica carinata variety derived from the plant of        embodiment 2 comprises at least a portion of the genetic makeup        of Brassica carinata variety DH-040.008.-   38. The method of embodiment 37, wherein steps (g) and (h) are    repeated until the Brassica carinata variety produced from variety    DH-040.008 has the desired trait and the physiological and/or    morphological characteristics set forth in one or more of Tables 1,    2, 3, 4, and 5, as determined at the 5% significance level, when    grown in the same location under the same environmental conditions    as variety DH-040.008.-   39. The method any one of embodiments 27 to 38, wherein the desired    trait is selected from the group consisting of male sterility and/or    fertility restoration, disease resistance, fungal resistance, pest    resistance, herbicide tolerance, abiotic stress tolerance, and    altered metabolism.-   40. The method of embodiment 39, wherein the desired trait is    herbicide tolerance and the herbicide is selected from, but not    limited to, the group consisting of glyphosate, glufosinate,    imidazolinones, and auxin analogues such as 2,4-D and dicamba.-   41. A plant, plant part, cell, or seed of a progeny Brassica    carinata variety produced by the method of any one of embodiments 27    to 40.-   42. A method for producing a Brassica carinata DH-040.008 CMS line    expressing a cytoplasmic male sterility (CMS) trait, comprising the    following steps:    -   (a) using a plant of Brassica carinata variety DH-040.008 as a        pollen donor for crossing with an existing Brassica CMS plant,        preferably a Brassica napus CMS variety or a Brassica juncea CMS        variety, to produce viable F₁ CMS plants;    -   (b) backcrossing the F₁ CMS plants with Brassica carinata        variety DH-040.008, using Brassica carinata DH-040.008 as a        pollen donor, to recover viable CMS BC₁ plants;    -   (c) repeatedly backcrossing CMS BC_(x) plants with Brassica        carinata variety DH-040.008, using Brassica carinata variety        DH-040.008 as a pollen donor, for up to 6 generations and        recovering viable CMS BC₆ plants (referred to as Brassica        carinata variety DH-040.008 CMS); and    -   (d) maintaining the Brassica carinata DH-040.008 CMS line by        crossing with Brassica carinata DH-040.008 wildtype (used as B        line) and harvesting of CMS seed from Brassica carinata        DH-040.008 CMS parent.-   43. A method for producing a Brassica carinata DH-040.008 Rf line    expressing a fertility restorer function gene (Rf) comprising:    -   (a) use of a plant of Brassica carinata variety DH-040.008 in        reciprocal crosses with an existing Brassica Rf plant,        preferably a Brassica carinata Rf, whereby viable F₁ progeny        plants are subsequently recovered then screened for presence of        Rf gene marker and whereby Rf F₁ plants are selected;    -   (b) backcrossing the Rf F₁ plants with Brassica carinata variety        DH-040.008, whereby viable BC₁ plants are subsequently recovered        then screened for presence of Rf gene marker and whereby Rf BC₁        plants are selected.    -   (c) repeated backcrossing of Rf BC_(x) plants with Brassica        carinata variety DH-040.008, for up to 6 generations as        described above, whereby, viable Rf BC₆ plants are ultimately        recovered (referred to as Brassica carinata variety DH-040.008        Rf); and    -   (d) maintenance of the Brassica carinata DH-040.008 Rf line by        self-crossing and harvesting of Rf seed from Brassica carinata        DH-040.008 Rf selfed parent.-   44. A method for producing hybrid seed from crossing a female parent    and a male parent, the method comprising the steps of:    -   (a) crossing a plant of variety DH-040.008 with another Brassica        carinata variety;    -   (b) growing the resultant F1 hybrid seed and selecting one or        more progeny plants that have a desired trait;    -   (c) recovering the hybrid seed, wherein the female parent is        Brassica carinata variety DH-040.008 CMS.-   45. A method for producing hybrid seed from crossing a female parent    and a male parent, the method comprising the steps of:    -   (a) crossing a plant of variety DH-040.008 with another Brassica        carinata variety;    -   (b) growing the resultant F1 hybrid seed and selecting one or        more progeny plants that have a desired trait;    -   (c) recovering the hybrid seed,    -   wherein the male parent is Brassica carinata variety DH-040.008        CMS.-   46. Hybrid seed produced from the method of any one of embodiments    42 to 45.-   47. Crushed, non-viable seed of Brassica carinata variety    DH-040.008, wherein a representative sample of the seed has been    deposited under NCIMB Accession number 42999.-   48. A cell of a seed, plant or plant part of Brassica carinata    variety designated DH-040.008, wherein a representative sample of    the seed has been deposited under NCIMB Accession number 42999.-   49. A protoplast of the cell of embodiment 48.-   50. A cell of a Brassica carinata plant or plant part having the    physiological and/or morphological characteristics set forth in one    or more of Tables 1, 2, 3, 4, and 5, as determined at the 5%    significance level, when grown in the same location under the same    environmental conditions as variety DH-040.008.-   51. The cell of embodiment 50, wherein the plant part is an ovule, a    leaf, pollen, a seed, an embryo a root, a root tip, a pod, a flower,    or a stalk.-   52. A tissue culture of protoplasts or regenerable cells of the cell    of embodiment 50 or 51.-   53. The tissue culture according to embodiment 52, wherein the    protoplasts or regenerable cells are produced from a tissue selected    from the group consisting of leaves, pollen, embryos, roots, root    tips, pods, flowers, ovules, and stalks.-   54. A Brassica carinata plant regenerated from the tissue culture of    embodiment 53, wherein the plant has the physiological and/or    morphological characteristics set forth in one or more of Tables 1,    2, 3, 4, and 5, as determined at the 5% significance level, when    grown in the same location under the same environmental conditions    as variety DH-040.008.-   55. A cell of a Brassica carinata plant regenerated from the tissue    culture of embodiment 53, wherein the plant has the physiological    and/or morphological characteristics set forth in one or more of    Tables 1, 2, 3, 4, and 5, as determined at the 5% significance    level, when grown in the same location under the same environmental    conditions as variety DH-040.008.-   56. Use of a plant of Brassica carinata variety DH-040.008 to    produce seed, wherein the seed is produced by self-crossing or open    pollination.-   57. Use of a plant of Brassica carinata variety DH-040.008 to    produce an F1 hybrid Brassica carinata seed, wherein the plant of    Brassica carinata variety DH-040.008 is either a female parent or a    male parent in a cross-fertilization.-   58. A cell of an F1 hybrid plant grown from the F1 hybrid seed    produced by the use of embodiment 57.-   59. A cell of an F1 hybrid plant grown from F1 hybrid seed produced    by a method comprising crossing a plant of Brassica carinata variety    DH-040.008 with a different Brassica carinata plant and harvesting    the resultant F1 hybrid carinata seed.-   60. Use of a plant of Brassica carinata variety DH-040.008 to    produce a Double Haploid variety.-   61. Use of embodiment 60, wherein the Double Haploid variety is    produced by a method comprising chromosome doubling introduced by    chemical or physical means.-   62. A cell of a Double Haploid (DH) variety produced from Brassica    carinata variety DH-040.008.-   63. Use of a plant of Brassica carinata variety DH-040.008 to    produce a Brassica carinata variety comprising a desired trait.-   64. Use of a plant of Brassica carinata variety DH-040.008 to    produce a Brassica carinata variety comprising a desired trait,    wherein the desired trait is conferred by a nucleic acid construct.-   65. The use of embodiment 64, wherein the nucleic acid construct is    introduced using polyethylene glycol (PEG) mediated uptake,    electroporation, ballistic infiltration using nucleic acid-coated    microprojectiles (gene gun), an Agrobacterium infiltration-based    vector, or a plant virus-based vector.-   66. The use of embodiment 64 or 65, wherein the nucleic acid    construct comprises a transgene, an RNAi construct, or an antisense    RNA construct.-   67. Use of a plant of Brassica carinata variety DH-040.008 to    produce a Brassica carinata variety comprising a desired trait,    wherein the desired trait is introduced by exposing seedlings or    microspores to a mutagenic agent.-   68. The use of embodiment 67, wherein the mutagenic agent is ethyl    methanesulfonate, N-ethyl-N-nitrosourea, or x-ray, gamma or    ultraviolet radiation.-   69. Use of a plant of Brassica carinata variety DH-040.008 to    produce a Brassica carinata variety comprising a desired trait,    wherein the desired trait is introduced by crossing a plant of    variety DH-040.008 with a plant of another Brassicaceae species    comprising the desired trait.-   70. The use of embodiment 69, wherein the Brassica carinata variety    produced from variety DH-040.008 has the desired trait and the    physiological and/or morphological characteristics set forth in one    or more of Tables 1, 2, 3, 4, and 5, as determined at the 5%    significance level, when grown in the same location under the same    environmental conditions as variety DH-040.008.-   71. The use of any one of embodiments 63 to 70, wherein the desired    trait is selected from the group consisting of male sterility,    disease resistance, fungal resistance, pest resistance, herbicide    tolerance, abiotic stress tolerance, and altered metabolism.-   72. The use of embodiment 71 wherein the desired trait is herbicide    tolerance and the herbicide selected from, but not limited to, the    group consisting of glyphosate, glufosinate, imidazolinones, and    auxin analogues such as 2,4-D and dicamba.-   73. A cell of a plant of a Brassica carinata variety comprising a    desired trait, wherein the Brassica carinata variety is produced    from Brassica carinata variety DH-040.008 by a method comprising the    steps of:    -   (a) crossing a plant of Brassica carinata variety DH-040.008        with another Brassica carinata variety comprising the desired        trait;    -   (b) growing the resultant F1 hybrid seed and selecting one or        more progeny plants that have the desired trait;    -   (c) backcrossing the selected progeny plants that have the        desired trait with plants of Brassica carinata variety        DH-040.008 to produce backcross progeny plants; and    -   (d) growing seed from the resultant backcross progeny plants and        selecting backcross progeny plants that have the desired trait        and at least a portion of the genetic make up of Brassica        carinata variety DH-040.008.-   74. The cell of embodiment 73, wherein the method to produce the    Brassica carinata variety further comprises repeating steps (c)    and (d) until the Brassica carinata variety has the desired trait    and the physiological and/or morphological characteristics set forth    in one or more of Tables 1, 2, 3, 4, and 5, as determined at the 5%    significance level, when grown in the same location under the same    environmental conditions as variety DH-040.008.-   75. A cell of a plant of a Brassica carinata variety produced from    variety DH-040.008, wherein the Brassica carinata variety comprises    a desired trait, and wherein the Brassica carinata variety was    produced by a method comprising introducing a nucleic acid construct    conferring the desired trait into a plant, plant part, or cell of    variety DH-040.008.-   76. The cell of embodiment 75, wherein the nucleic acid construct is    introduced using polyethylene glycol (PEG) mediated uptake,    electroporation, ballistic infiltration using nucleic acid coated    microprojectiles (gene gun), an Agrobacterium infiltration-based    vector, or a plant virus-based vector.-   77. The cell of embodiment 75 or 76, wherein the nucleic acid    construct comprises a transgene, an RNAi construct, or an antisense    RNA construct.-   78. The cell of any one of embodiments 75 to 77, wherein the    Brassica carinata variety produced from variety DH-040.008 comprises    the desired trait and the physiological and/or morphological    characteristics set forth in one or more of Tables 1, 2, 3, 4, and    5, as determined at the 5% significance level, when grown in the    same location under the same environmental conditions as variety    DH-040.008.-   79. A cell of a plant of a Brassica carinata variety produced from    variety DH-040.008, wherein the Brassica carinata variety comprises    a desired trait, and wherein the Brassica carinata variety was    produced by a method comprising the steps of:    -   (a) crossing a plant of variety DH-040.008 with a plant of        another Brassica carinata variety comprising the desired trait;    -   (b) growing the resultant F1 hybrid seed and selecting one or        more progeny plants that have the desired trait;    -   (c) self-pollinating the progeny plants that have the desired        trait to produce further progeny plants;    -   (d) growing the resultant further progeny plants and selecting        further progeny plants that have the desired trait and at least        a portion of the genetic make up of Brassica carinata variety        DH-040.008.-   80. The cell of embodiment 79, wherein steps (c) and (d) are    repeated until the Brassica carinata variety produced from variety    DH-040.008 has the desired trait and the physiological and/or    morphological characteristics set forth in one or more of Tables 1,    2, 3, 4, and 5, as determined at the 5% significance level, when    grown in the same location under the same environmental conditions    as variety DH-040.008.-   81. A cell of a plant of a Brassica carinata variety produced from    variety DH-040.008, wherein the Brassica carinata variety comprises    a desired trait, and wherein the Brassica carinata variety was    produced by a method comprising the steps of:    -   (a) crossing a plant of variety DH-040.008 with a plant of        another Brassica carinata variety comprising the desired trait;    -   (b) growing the resultant F1 hybrid seed and selecting one or        more progeny plants that have the desired trait;    -   (c) backcrossing the selected progeny plants that have the        desired trait with plants of from the Brassica carinata plant of        embodiment 2 to produce backcross progeny seed; and    -   (d) growing the resultant backcross progeny seed and selecting        backcross progeny plants that have the desired trait and at        least a portion of the genetic make up of Brassica carinata        variety DH-040.008.-   82. The cell of embodiment 81, wherein steps (c) and (d) are    repeated until the Brassica carinata variety produced from variety    DH-040.008 has the desired trait and the physiological and/or    morphological characteristics set forth in one or more of Tables 1,    2, 3, 4, and 5, as determined at the 5% significance level, when    grown in the same location under the same environmental conditions    as variety DH-040.008.-   83. A cell of a plant of a Brassica carinata variety produced from    variety DH-040.008, wherein the Brassica carinata variety comprises    a new trait and is produced by a method comprising exposing    seedlings or microspores to a mutagenic agent and allowing the    surviving fraction to develop into mature plants.-   84. The cell of embodiment 83, wherein the mutagenic agent is ethyl    methanesulfonate, N-ethyl-N-nitrosourea, or x-ray, gamma or    ultraviolet radiation.-   85. A cell of a plant of a Brassica carinata variety comprising a    desired trait, wherein the Brassica carinata variety is produced by    a method comprising:    -   (a) crossing a plant of variety DH-040.008 with a plant of        another Brassicaceae species comprising the desired trait;    -   (b) producing F1 plants using embryo rescue techniques to        recover viable F1 plants from the cross or growing F1 seeds;    -   (c) self-pollinating the F1 plants that have the desired trait        and carinata character;    -   (d) producing F2 plants using embryo rescue techniques to        recover viable F1 plants from the cross or growing F2 seeds;    -   (e) self-pollinating the F2 plants that have the desired trait        and carinata character;    -   (f) producing progeny plants using embryo rescue techniques to        recover viable F3 plants or growing F3 seeds;    -   (g) self-pollinating the progeny plants that have the desired        trait and carinata character to produce further progeny plants;        and    -   (h) selecting the progeny plants with the desired trait and        carinata character.-   86. The cell of embodiment 85, wherein steps (g) and (h) are    repeated until the Brassica carinata variety produced from variety    DH-040.008 has the desired trait and the physiological and/or    morphological characteristics set forth in one or more of Tables 1,    2, 3, 4, and 5, as determined at the 5% significance level, when    grown in the same location under the same environmental conditions    as variety DH-040.008.-   87. The cell of any one of embodiments 73 to 86, wherein the desired    trait is selected from the group consisting of male sterility,    disease resistance, fungal resistance, pest resistance, herbicide    tolerance, abiotic stress tolerance, and altered metabolism.-   88. The cell of embodiment 87, wherein the desired trait is    herbicide tolerance and the herbicide is selected from, but not    limited to, the group consisting of glyphosate, glufosinate,    imidazolinones, and auxin analogues such as 2,4-D and dicamba.-   89. Use of a plant of Brassica carinata variety DH-040.008 to    produce a commercial crop.-   90. Use of a plant of Brassica carinata variety DH-040.008 to    produce a commercial plant product.-   91. The use of embodiment 90, wherein the commercial plant product    comprises oil, meal, protein isolate, or biofumigant.-   92. A method for producing a group of cultivated plants of Brassica    carinata variety DH-040.008 in a field, wherein harmful    microorganisms are controlled by the application of a composition    comprising one or more microbiocidal ingredient.-   93. The method of embodiment 92, wherein the one or more    microbiocidal ingredient is selected from the group comprising    iprodione, prothiaconazole vinclozolin, boscalid, carbathiin,    thiram, difenoconazole, metalaxyl, sedaxane, fludioxonil, penflufen,    trifloxystrobin, and sedaxane.-   94. The method of embodiment 93, wherein the composition comprising    one or more microbiocidal ingredient is applied as a seed treatment    or seed coating.-   95. A method for producing a group of cultivated plants of Brassica    carinata variety DH-040.008 in a field, wherein insect pests are    controlled by the application of a composition comprising one or    more insecticidal ingredient.-   96. The method of embodiment 95, wherein the insecticidal ingredient    is applied as a seed treatment or foliar treatment.-   97. A method for cleaning seed of Brassica carinata variety    DH-040.008, or seed from a plant produced from Brassica carinata    variety DH-040.008, to remove foreign material from the surface of    the seed.-   98. A method for cleaning seed of Brassica carinata variety    DH-040.008, or seed from a plant produced from Brassica carinata    variety DH-040.008, to remove any debris or low quality, infested,    or infected seeds, or seeds of different species.

Definitions

In the following description and tables, many terms are used. To aid ina clear and consistent understanding of the specification, the followingdefinitions and evaluation criteria are provided.

Abiotic stress is defined as the negative impact of non-living factorson the living organisms in a specific environment. Examples of abioticstress include, but are not limited to, drought, water-logging orflooding, extreme temperatures, extreme salinity, and mineral toxicity.

Agronomic practice refers to any of a set of cultivation practices ortechniques that attempt to maximize the health and productivity of acrop. The agronomic practices used are an important factor ininterpreting results from field studies of various kinds.

Allele refers to one or more alternative forms of a gene locus thatrelate to one trait. Diploid organisms, i.e., organisms with two sets ofchromosomes, have one copy of each gene (and therefore, one allele) oneach chromosome. If both the alleles are the same, they are homozygous.If the alleles are different, they are heterozygous.

Alternaria resistance is typically rated on a scale from 0-5; 0=nosymptoms were observed, 1=indicates infection of pods only, 2=diseaseprevalence on 25% of upper plant, 3=50% prevalence, 4=75% prevalence,and 5=disease symptoms are seen throughout entire plant and consideredsevere

Average refers to the arithmetic mean. “Substantially equivalent” or“statistically equivalent” refers to a characteristic that, whencompared, does not show a statistically significant difference from themean. In contrast, “statistically different” or “statisticalsignificance” refers to a characteristic that, when compared, showsstatistically significant differences from means of the samecharacteristic of another group or groups. Most often, statisticalsignificance of differences is measured at levels of P<0.05 usingstandard tests to compare Least Square Means, such as Tukey's HSD orStudent's t-test.

Backcrossing means a traditional breeding technique used to introduce atrait to a plant line or variant from a donor plant to a recurrentplant. An initial cross is made between the donor and recurrent parentplants to produce progeny plants. Progeny plants having the desiredtrait are then crossed to the recurrent parent. This process ofbackcrossing is repeated for several breeding cycles until the progenyplants are indistinguishable from the recurrent parent, except for thetrait from the donor parent.

Breeding line (or Plant line) refers to a unique, reproducible carinatatype, and is distinguishable from other carinata types based on itsgenotype and phenotype. Most often, a breeding line refers to a typethat is mostly or completely homozygous, such as a DH line or a highlyinbred line (five or more generations inbred).

Carinata refers to seeds or plants of the species Brassica carinatacontaining both the B genome from Brassica nigra and the C genome fromBrassica oleracea (Nagahuru, 1935). The terms “Brassica carinata varietyDH-040.008”, “carinata variety DH-040.008”, “variety DH-040.008”, and“DH-040.008” are used interchangeably herein and refer to a plant ofBrassica carinata variety DH-040.008, representative seed of whichhaving been deposited under NCIMB Accession number 42999.

Check variety (or Check line) are carinata genotypes considered to bethe standard for overall performance, or for a specified trait, in agiven growing region. Often, the current commercial varieties are usedas check lines for the regions in which they are grown, and this is thestandard against which new potential varieties are tested. Examples ofuseful commercial check varieties include, but are not limited to,Brassica carinata varieties AAC-A120 and AGRO44-312D-HP11(WO2017/181276A1; henceforth referred to as “AG044-HP11” or “HP11”)

Days to Flowering (initiation of flowering) refers to the number of daysfrom planting until 50% of the plants in a planted area have at leastone open flower.

Depth of canopy, measured in cm, is the distance from the firstincidence of pods on a plant (average within a plot) to the top of thepod canopy.

Diploid refers to cell or a plant with two sets of chromosomes. One setcomes from each parent.

Double Haploid, Doubled Haploid, Doubled Haploidy (DH) refers to ahaploid cell or plant that has undergone a doubling of its chromosomesto produce a functional diploid.

Duration of flowering is the number of days between the initiation offlowering and end of flowering.

Early (season) vigour is a rating, usually on a scale of 1 to 7, thatreflects how a variety develops in its early stages (about 4 weeks afterseed) in terms of putting on leaf area and competing with weeds.Typically, experimental varieties or lines are compared with a checkvariety or line, which is rated as a “4”. A new variety with slightlymore leaf cover and advanced development would be rated a “5”, or moresignificantly a “6”, and so forth. 1=significantly less than checkvariety; 7=significantly greater uniformity than check variety.

End of flowering is the number of days from planting to whenapproximately 90% of flowers have lost their petals.

Erucic Acid content is the weight percentage (wt %) of fatty acids inthe form of C22:1 among all major fatty acid types found in carinata.Most often, this is estimated by Near Infrared Spectroscopy (NIR) ofmature seeds at less than 6% moisture. The NIR is calibrated using alarge array of samples whose fatty acid profile is determined byAmerican Oil Chemists Society (AOCS) Official Method Ce1-66 Fatty AcidComposition by Gas Chromatography. This is one of the official methodsrecommended by the Western Canada Canola/Rapeseed Recommending Committee(WCC/RCC).

Extent of branching refers to the number of pod-bearing secondarybranches (racemes) off the main stem. This is often estimated using anaverage of at least eight plants per breeding line.

Embryo Rescue techniques refers to in vitro techniques whose purpose isto promote the development of an immature or weak embryo into a viableplant. This methodology is commonly used to rescue viable embryos fromcrosses between different but closely related species, i.e.,interspecific crosses.

Fatty acid content means the typical percentages by weight of fattyacids present in the endogenously formed oil of the mature whole driedseeds, as determined by Near Infrared Spectroscopy at less than 6% seedmoisture. The NIR is calibrated using a large array of samples whosefatty acid profile is determined by American Oil Chemists Society (AOCS)Official Method Ce1-66 Fatty Acid Composition by Gas Chromatography.This is one of the official methods recommended by the Western CanadaCanola/Rapeseed Recommending Committee (WCC/RCC). Aside from erucic acid(C22:1), the most significantly occurring fatty acids in carinata areoleic acid (C18:1), linoleic acid (C18:2), linolenic acid (C18:3), andeicosenoic acid (C20:1).

Flower Petal Colouration means the colouration of open exposed petals onthe first day that flowering is observed. In carinata, varieties aremost often categorized as either W=White or Y=Yellow. Occasionally, forvarieties with a very light yellow colour, a third category will beemployed: PY=Pale Yellow.

Frost tolerance means the ability of young plants to withstand frosts inareas where carinata is grown during the winter season. These frosts aremost likely to occur from the five-leaf to early bolting plantdevelopment stages, depending on time of planting. This is typicallymeasured 3-5 days following a hard frost event and rated using a scaleof visual percentage of injury (leaf bleaching and death); where “0” isno injury and “10” is 100% plant damage, and each successive numberindicates an additional 10% injury to leaf and stem tissue.

Gene silencing means the interruption or suppression of the expressionof a gene at the level of transcription or translation.

Genotype refers to the genetics or DNA sequence of individual carinatalines, as opposed to their actual appearance, which is called thephenotype.

Glucosinolate (GSL) content means the total glucosinolate content ofmature seed at less than 6% moisture, given in units of μmol/g, asanalyzed using Near Infrared Spectroscopy. In carinata, the majority ofglucosinolate content is in the form of sinigrin (chemical name:allylglucosinolate or 2-propenylglucosinolate). Most often, the totalglucosinolate content of seeds is estimated by Near InfraredSpectroscopy (NIR) of mature seeds at less than 6% moisture. The NIR iscalibrated using a large array of samples with known GSL contentdetermined previously by one or more methods known in the art including,but not limited to, gas chromatography of TMS-derivatives, and HPLC ofdesulfoglucosinolates.

Grain means the seed produced by carinata crops that are intended forprocessing for oil or feed uses. This is in contrast with parent seed orplanting seed, which is intended for growth of another generation ofplants.

Growing Degree Days refers to the accumulation of heat units above abase temperature over time and is often strongly correlated with rate ofplant development. The daily GDD units are calculated by subtracting theaverage temperature (° C.) (average of daily maximum and minimumtemperature) from the baseline of 5° C. These units accumulate beginningthe day after planting.

Growing Environment refers to a particular area (typically grouped bygeography) or controlled environment where plants are grown andevaluated. Expression of various characteristics or traits, such asplant height or days to maturity, can be greatly influenced by theenvironment in which it is grown. This is also commonly known asGenotype x Environment interaction, or G x E.

Haploid refers to a plant, or a cell of a plant, that has only one setof chromosomes. Haploid plants can be produced artificially, and theirsingle set of chromosomes can be doubled to produce a doubled haploid.

Height of first pod, measured in cm, refers to the distance from groundlevel to the first incidence of pods on a plant (average within a plot).

Herbicide Resistance means the resistance to various herbicides whenthat herbicide is applied at standard recommended application rates andtiming; and is expressed on a scale of 0 to 10; where “0” is nosymptoms, and a rating of “10” means entire plant is brown and curled.Each successive number indicates approximately 10% more damage or effectof herbicide on the plant: a rating of “2” indicates several leaves withslight yellowing and curl, “4” is several yellowing or dying leaves andmore noticeable curl, “6” is a greater number and severity of yellowing,curling, and dead leaves, and “8” indicates that most leaves arecompletely yellow or dead.

Hybrid variety or F1 hybrid refers to the seeds harvested from crossingtwo inbred (nearly homozygous) parental lines. This is most oftenaccomplished in carinata through the development of inbred parents usingthe Ogura pollination control system, where one parent is functionallymale sterile (CMS or A-line) and the other restores fertility to the F1hybrid (Rf or R-line).

Leaf colour means the colour of the leaf blade of lower leaves at thelate bolting to early flowering stages (on a plot basis). A categoricalscale is most often used, where DG=Dark green; G=Green; LG=Light green;BG=Bluish green.

Leaf glaucosity refers to the presence or absence of a fine whitishpowdery coating on the surface of the leaves, and the degree thereof,when present. Numerical scale used to rate this trait: 3=weak, 5=medium,7=strong.

Leaf Length is the total length of the leaf from attachment of thepetiole to the stem, to the tip of the blade, measured in cm, of lowerleaves at the late bolting to early flowering stages. It is estimatedusing an average of measurements from at least 30 plants.

Leaf number of Lobes means the frequency of leaf lobes, when present, oflower leaves at the late bolting to early flowering stages. 3=few,5=medium, 7=many.

Leaf Width is the width, in cm, of the widest part of the leaf blade oflower leaves at the late bolting to early flowering stages. It isestimated using an average of measurements from at least 30 plants.

Locus refers to a specific location in a chromosome.

Lodging, is the displacement of stems from their vertical and properplacement, observed as extent to which plants of a given variety remainupright; upright plants are easier to harvest with harvesting equipment.In some trials, lodging is measured on a scale of 1-5, where1=significantly less lodging than check variety; 5=significantly greaterlodging than check variety. In other trials, lodging is measured on ascale of 1-7, where 1=plot flat on the ground; 2=most of plot on ground;3=75% lodged; 4=50% lodged; 5=25% lodged; 6=only slight lodging/stembending noticed; 7=no lodging.

Maturity may be determined based on Seed Maturity, PhysiologicalMaturity or both.

Near Infrared Spectroscopy (NIR) refers to a non-destructive,spectrometric method commonly used to assess seed quality parameters ingrain producing crops. Estimates of total seed oil content and fattyacid profile, as well as total glucosinolate and protein content on awhole seed basis, are examples of parameters for which NIR is commonlyemployed.

Number of seed-bearing pods is determined by counting the number of podscontaining at least one viable seed on the main raceme, as measured inat least eight plants of the same line at full pod set stage.

Oil content means the typical percentage by weight (wt %) of oil presentin the mature whole dried seeds, at less than 6% moisture, as determinednear infrared (NIR) spectroscopy (AOCS Procedure Am 1-92 Determinationof Oil, Moisture and Volatile Matter, and Protein by Near-InfraredReflectance).

Open-pollinated (OP) variety refers to varieties that are maintained andpropagated primarily by means of self-pollination, and for which thenext generation will be largely equivalent to the previous.

Pedicel Length is the typical length of the silique stem observed whenmature, measured in mm, for 30 pods taken per plot, from pods occurringon mid-third of the main raceme.

Petiole Length is the length of the petiole, measured in cm, fromattachment of the petiole to the stem, to the start of the leaf blade,as observed in lower leaves at the late bolting to early floweringstages.

Phenotype refers to the outward appearance or manifestation of giventraits of varieties, individual plants, or plant parts (such as leavesor seeds).

Physiological maturity means the number of days from planting to thestage when pods with seed change colour, from green to a dry tan lookand when about 70-80% of stems and pods dry-down, corresponding tostages 90 and beyond of the BBCH scale (Meier, et al., 2009).

Plant includes the whole plant, or any plant parts such as plant organs,plant cells, plant protoplasts, plant cell cultures, or plant tissuecultures form which whole plants can be regenerated, plant callus, plantcell clumps, plant transplants, seedlings, plant cells that are intactin plants, plant clones or micropropagations.

Plant height means the overall plant height from ground level to theaverage of the top of the plant canopy, in cm, taken betweenmid-flowering and full pod-set stages of plant development.

Plant length is the length of the plant, in cm, measured at maturity.For this trait, it is typically an average 30 or more plants measuredindividually from the same variety.

Plant part includes, but is not limited to, harvested tissues, fruits,organs, plant cuttings, vegetative propagations, embryos, pollen,ovules, flowers, leaves, fruits, fruit flesh, seeds, clonally propagatedplants, roots, stems, stalks, root tips, grafts, parts of any of theseand the like, or derivatives thereof, preferably having the same geneticmakeup (or very similar genetic make up) as the plant from which it isobtained. Plant part also includes any developmental stage, such asseedlings, cutting prior or after rooting, mature and/or immatureplants, or mature and/or immature leaves.

Ploidy refers to the number of sets of chromosomes exhibited by theline, for example, diploid (two sets) or tetraploid (four sets).

Plot refers to a group of carinata plants, grown in one or multiple rowsof varying lengths, which is often used as the basic unit of measurementfor a number of traits. These are often, but not always, associated withreplicated yield trials.

Pod shatter loss is the amount of seed lost from shattered pods atharvest; seeds are collected in three 7″×13″ pans placed 3′ from eitherend or at center of each plot and converted to kg/ha.

Pod shatter resistance, or shatter resistance, means the resistance tosilique shattering observed at seed maturity. 1=not tested, 3=poor,5=fair, 7=good, 9—does not shatter.

Pod (silique) Beak length, measured in mm, for 30 pods taken per plot,from pods occurring on mid-third of the main raceme.

Pod (silique) length is the length of the pod not including the pedicelor beak, measured in mm, for 30 pods per plot taken from the mid-thirdof the main raceme.

Pod (silique) width is the typical pod width when mature, for 30 podsper plot taken from the mid-third of the main raceme. Rating scale istypically categorical: 3=narrow; 5=medium, 7=broad.

Primary raceme length, measured in cm, means the measurement of the mainraceme from the last (youngest) branch to the top of the inflorescence.This measurement often taken from end flowering to full pod set stage ofplant development.

Progeny refers to plants derived from a plant of Brassica carinatavariety DH-040.008. Progeny may be derived by regeneration of cellculture or tissue culture of a plant of carinata variety DH-040.008,self-pollination of a plant designated DH-040.008, crossing at least oneplant of Brassica carinata variety DH-040.008 with a plant of anothervariety or line of Brassica carinata or other Brassicaceae species, orby producing seeds of a plant of Brassica carinata variety DH-040.008.

Protein content means the typical percentage by weight (wt %) of proteinin the oil-free meal of the mature whole dried seeds, at less than 6%moisture, analyzed using near infrared (NIR) spectroscopy (AOCSProcedure Am 1-92 Determination of Oil, Moisture and Volatile Matter,and Protein by Near-Infrared Reflectance).

Randomized Complete Block Design (RCBD) is the most common experimentaldesign used in standard replicated yield testing, typically consistingof four replications of various breeding lines or varieties, having thecomplete set of entries arranged in different randomizations for eachreplication.

Recovery from frost damage, is assessed by rating the same plots at acouple additional time points beyond the initial rating, often at twoand three weeks following a hard frost event.

Regeneration means the development of a plant from cell culture ortissue culture or vegetative propagation.

Replication refers to a series of ratings or observations, taken fromdifferent plots or samples of the same variety, breeding line, or traitthereof, from plants grown at the same location or within a given set oftrials. Replicated measurements are critical for greater accuracy instatistical analysis.

Resistance means the ability of a plant to withstand exposure to aninsect, disease, herbicide or other potentially stress-inducingcondition. A resistant plant variety or hybrid will have a level ofresistance higher than a comparable check variety or hybrid.

Saturated Fatty Acids refer to Fatty Acids in which carbon chains arelinked by single bonds; they lack unsaturated links between carbonatoms. The Saturated Fatty Acid content of oil in the seed is

Sclerotinia resistance is typically rated on a scale from 0-5; 0=nosymptoms were observed, 1=infection of pods only, 2=disease prevalenceon 25% of upper plant, 3=50% prevalence, 4=75% prevalence, and 5=diseasesymptoms are seen throughout entire plant and considered severe.

Seed colour (NIR), means the seed coat colour of typical mature seedsbased on NIR analysis. For this objective measurement, the colour of anaggregate sample of seed (1-5 g of seed) is determined by using the FOSXBR, or equivalent, near infrared spectrophotometer as a reflectancespectrophotometer over the visible range (400 nm to 700 nm) to providean objective description of the carinata seed color, essentially asdescribed (Black and Panozzo, 2004) except that seed color is describedin terms of the Hunter L*A*B* color space rather than CIEL*A*B* colorspace.

Seed colour (visual), is a subjective categorization of the predominantseed coat colour at seed maturity for a given variety or breeding line.These are generally grouped into the following categories: Y=Yellow orbright yellow; DY=Dark yellow; LB=Light brown; B=Brown; DB=Dark brown;O=Orange.

Seed Maturity refers to number of days from planting to the stage when70-80% seeds on main raceme had seeds with complete colour change andhard when pressed between fingers.

Seeds per pod, means the average number of seeds per pod, in at least 30pods taken from the mid-third of the main stalk.

Seed weight (thousand seed weight, or TKW), means the weight, in grams,of 1,000 typical seeds determined at maturity, as measured at a seedmoisture content of approximately 5-6%.

Self-pollination or “selfing” means the self-pollination of a plant bytransfer of pollen from an anther to a stigma of the same plant.Carinata is a self-compatible species, meaning there are no genetic orphysiological impediments to successful self-pollination.

Stand is the number of plants counted in 4 rows and converted toplants/m². Recorded 2-4 days before harvest.

Stem colour refers to the predominant stem colour at bolting stage,depending on levels of anthocyanin manifest in the stem. This isgenerally categorical, characterized as G=Green; LP=Light Purple; andP=Purple or dark purple.

Tissue culture refers to a composition comprising isolated cells of thesame or a different type or a collection of such cells organized intopart of a plant. This technique is an integral part of breedingtechniques such as doubled haploidy, whereby new diploid plants of aunique, homozygous genetic composition are generated in lab conditionsto the point where normal plant growth can occur in the field or in agreenhouse.

Tolerance is commonly used in the context of plants affected by abioticstress, diseases, or pests and is used to describe an improved level offield resistance.

Traditional plant breeding techniques include, but are not limited to,crossing, selfing, selection, double haploid production, embryo rescue,marker assisted selection, mutation breeding, backcross breeding, singleseed descent, and any other method known to the breeder other thangenetic modification and transformation/transgenic methods, by which agenetically heritable trait can be transferred from one carinata line orvariety to another, or traits of interest fixed in one geneticbackground.

Trait introgression refers to plants within a variety have been modifiedin a manner that retains the overall genetics of the variety and furthercomprises one or more loci with a specific desired trait, such as malesterility, disease, or herbicide resistance. Backcrossing followed byinbreeding or DH line generation is a common methodology to achieve thisobjective. Using this technique, one or more value added traits may beintroduced into a single carinata variety.

Transgene means a genetic locus comprising a DNA sequence which has beenintroduced into the genome of a Brassica carinata plant bytransformation. A plant comprising a transgene stably integrated intoits genome is referred to as a transgenic plant.

DETAILED DESCRIPTION

In one aspect, the present invention relates to cells, seeds, plants,and plant parts of Brassica carinata variety DH-040.008, alternatively“carinata variety DH-040.008”, “variety DH-040.008”, or “DH-040.008”,for which a representative sample of the seed has been deposited underNCIMB Accession number 42999.

In another aspect, the present invention relates to Brassica carinataplant or plant part produced or derived from seeds, plants, and plantparts of Brassica carinata variety DH-040.008, as well as to all progenyof Brassica carinata variety DH-040.008 produced by one or morebreeding, mutagenesis, tissue culture, or genetic modificationtechniques and having essentially all the physiological andmorphological characteristics of a plant or plant part of Brassicacarinata variety DH-040.008 when grown in the same location under thesame environmental conditions as variety DH-040.008.

In another aspect, the present invention relates to Brassica carinataplant or plant part produced or derived from seeds, plants, and plantparts of Brassica carinata variety DH-040.008, as well as to all progenyof Brassica carinata variety DH-040.008 produced by one or morebreeding, mutagenesis, tissue culture, or genetic modificationtechniques and having the physiological and/or morphologicalcharacteristics set forth in one or more of Tables 1, 2, 3, 4, and 5, asdetermined at the 5% significance level, when grown in the same locationunder the same environmental conditions as Brassica carinata varietyDH-040.008.

The present invention also relates to any plants produced or derivedfrom seeds, plants, and plant parts of Brassica carinata varietyDH-040.008, that exhibit “carinata character”.

For the purpose of the present invention, plants exhibiting “carinatacharacter” have an erect, upright bearing, are highly branching, withwell-developed and aggressive tap root systems (Barro and Martin, 1999).Leaves are generally wide elliptic in shape with weak-medium dentation,medium glaucosity, and very sparse pubescence. Seeds are globose, 1-1.5mm in diameter and finely reticulated (Mnzava and Schippers, 2004) andvary from yellow to yellow-brown to brown in colour (Getinet 1987;Rahman and Tahir, 2010).

Plants exhibiting “carinata character” may be produced from the seed ofBrassica carinata variety DH-040.008; a representative sample of theseed has been deposited under NCIMB Accession number 42999 or from anycell, plant or plant part produced from the seed of Brassica carinatavariety DH-040.008, using one or more (conventional) plant breedingtechnologies, cell culture or tissue culture technologies, and/ortransgenic technologies.

Plant Breeding Technologies

Critical to the development and breeding of any crop is the ability tomake use of genotypic and phenotypic diversity. Breeding strategies makeuse of the plant's method of pollination: self-pollination, where thepollen from one flower is transferred to the same or another flower onthe same plant or a genetically identical plant; sib-pollination, whenindividuals with the same family or line are used for pollination; orcross-pollination, where the pollen comes from a flower on a geneticallydifferent plant from a different family or line.

For practical application, a breeder initially selects and crosses twoor more parental lines, followed by repeated selfing and selection toproduce many unique genetic combinations. For a newly developing cropsuch as carinata, it is necessary to be able to obtain a sufficient poolof genetic material to identify genetic backgrounds more adapted totarget geographies (i.e. a better starting point), as well as variationfor traits of interest. This allows for crossing or other modificationsto be done to identify genetic combinations superior to the typesalready tested. Thus, an important initial objective is the collectionand characterization of as large of a collection of genetic backgroundsas possible, for each target geography. Parental lines are selectedbased on breeding priorities and the unique combination of traitsavailable in potential crossing parents. The selection of parents ofthese crosses is critical to the effectiveness of a breeding program.Parental lines may be closely or distantly related lines of a singleplant species or may be two different species of the same genus.

Crossing, of (inbred) parental lines, by sexual hybridization, istypically done manually in controlled conditions. Often, two or threerounds of crossing are needed to accumulate beneficial alleles into asingle genetic background. This includes evaluating offspring of across, selecting the most desirable (inbred) lines as future parents,and making the next round of parental selection based on prioritytargets. Theoretically, billions of different genetic combinations canbe produced through a combination of mutagenesis, selfing, and crossing.Since the breeder has no direct control at the cellular level, twobreeders will never independently develop the same variety of carinataplants have the same traits.

In each cycle of selection and evaluation, the breeder selects germplasmto advance to the next generation by growing individual plants in thechosen geography, soil and climate conditions and collecting phenotypicdata reflective of actual performance that would be realized by a seedproducer. Traits collected focus on those that would be of agronomic oreconomic benefit in the crop. Examples of traits characterized in acarinata breeding program include, but are not limited to, early plantvigor, plant height, branching habit, days to flower, silique density,flower petal color, pod size, reaction to heat and water stress, diseasesusceptibility, and pod shatter tolerance.

In a typical carinata breeding program, the breeder initially selectsand crosses two or more parental Brassica carinata lines, followed byrepeated selfing and selection to produce many unique geneticcombinations, which are evaluated for overall agronomic potential aswell as specific traits. Such recurrent selection can be used to improvea population of either self- or cross-pollinating Brassica.Intercrossing of several parents creates a genetically variablepopulation of heterozygous individuals and the best plants are selectedbased on individual superiority, outstanding progeny and/or excellentcombining ability. New populations are created by further intercrossingand selection. This method is useful for the improvement ofquantitatively inherited traits controlled by numerous genes.

Backcrossing may be used to transfer genes for a simply inherited,highly heritable trait from a donor parent to the recurrent parent.After the initial cross, individual plants with the desired trait of thedonor parent are selected and backcrossed to the recurrent parent forseveral generations. The resulting progeny are expected to have theattributes of the recurrent parent and the desired trait from the donorparent. Backcrossing may be used in conjunction with pedigree breeding.Pedigree breeding and recurrent selection methods are used to developvarieties from breeding populations. Pedigree breeding starts with thecrossing of two genotypes, each with one or more desirablecharacteristics that is lacking in the other or that complement eachother. Additional genotypes can be included in the breeding populationif the original parents do not provide all the desired characteristics.In the pedigree method of breeding, five or more generations of selfingand selection may be used. For example, crossing of the two initialparents (the donor and recurrent parents) produces an initial F1population, from which an F2 population is produced by selfing one orseveral F1 plants or by intercrossing two F1 plants (sib mating).Selection of hybrids with desired combination of traits may be conductedwith the F2 population, or with the F3 or subsequent population.

Plants that have been self-pollinated and selected for severalgenerations become homozygous at nearly all gene loci and produce auniform population of breeding progeny or inbred lines. Subsequentcrosses with two different homozygous (inbred) lines produces a uniformpopulation of hybrid plants that may be heterozygous for many gene loci.Crossing two plants each heterozygous at a number of gene loci willproduce a population that differ genetically and will not be uniform.

Doubled haploid (or DH) technology allows for the generation ofcompletely homozygous lines, which are a combination of genes of theparental lines, in a single generation. This accelerates the process ofinbred line development dramatically as the process from seeding ofparental lines to obtaining seed for a resulting inbred population fromthose parents will generally take about 18 months. To achieve a highlyhomozygous line using traditional self-pollination generally will takefive or six growth cycles which, in the case of carinata, wouldrepresent three years or six growth cycles, with two cycles completedper year. In DH technology, using appropriate in vitro conditions,haploid microspores from an F1 plant can be induced to differentiateinto diploid embryos and subsequently plantlets. This techniquetypically relies on a percentage of regenerated plants to undergospontaneous doubling (usually in the range of 20 to 60% of plants,depending on several factors), whereas the remaining plants will remainhaploid and sterile. To increase the efficiency of space used for seedincrease, such as in the greenhouse or field, a flow cytometer is usedto distinguish at an early stage the chromosome content “n” or “2n” ofeach plant. Thus, the sterile plants can be discarded at an early stage.

In some instances, a desirable trait may not reside within the speciesof interest, specifically Brassica carinata. In that case, it may bepossible to transfer the trait via interspecific or wide crossing. Forinterspecific crossing, one parent is Brassica carinata varietyDH-040.008 and the second parent may be a different species of Brassica,including but not limited to B. juncea (L). Czern. (brown mustard), B.napus L. (rape, Argentine canola), B. nigra (L.) W. D. J. Koch (blackmustard), B. rapa L. (field mustard, Polish canola), and B. oleracea L.(cabbage, broccoli, cauliflower, brussels sprouts, kohlrabi and kale).In other instances, the other Brassicaceae species including but notlimited to Brassica alba, Brassica hirta, Brassica juncea, Brassicanapus, Brassica nigra, Brassica oleracea, Brassica rapa, Sinapus alba,and Camelina sativa.

The methodology for performing interspecific crosses is similar to thatdescribed for within-species crosses described above. However, unlikeintraspecific crosses, the likelihood that the resulting progeny willproduce viable seed is very low and thus represents a formidablechallenge to the success of this technique. To overcome this potentialblock, embryo rescue techniques are often employed to recover viableoffspring from the cross. Essentially, this relies on the progenysurviving until the embryogenic stage at which point it can be dissectedfrom the silique and placed into artificial growth medium. Underappropriate conditions, the cultured embryo can survive and be inducedto differentiate into a plantlet, which can be grown into a matureplant. Successive rounds of embryo rescue may be needed until inbredprogeny, or backcross-derived progeny, are stable and can producefertile offspring without intervention. Often molecular markers, whereavailable, are used to trace a specific allele from a related speciesinto an adapted background in the target species, using repeated cyclesof backcrossing.

In addition, to minimize the relative proportion of the donor genomefrom the non-carinata species, several rounds of back-crossing of therescued plants with the Brassica carinata parent may be required togenerate progeny having carinata character and the desired trait, whichare stable and can produce fertile offspring.

Selection

Breeding nurseries are often the first cycle of evaluation of breedingpopulations. Generally, nurseries utilize single or paired rows withfrequent checks (i.e. the best available commercial germplasm for aspecified geography.

Various methods are used to screen breeding populations for individualplants with desired characteristics. Single seed descent procedurerefers to planting a segregating population, harvesting a sample of oneseed per plant, and using the one-seed sample to plant the nextgeneration. When the population has advanced from the F2 to the desiredlevel of inbreeding, the plants from which lines are derived will eachtrace to different F2 individuals. Since the number of plants in apopulation declines each generation due to failure of some seeds togerminate or produce seed, not all the F2 plants will be represented bya progeny in the final generation.

A multiple-seed procedure is when pods form each plant in a populationare harvested and threshed together to form a bulk. Part of the bulk isused to plant the next generation and part is put in reserve. Theadvantage of this method is that it is faster to harvest seed with amachine than to remove one seed from each plant by hand. If desired,Double(d) Haploid methods can be used to extract homogeneous(homozygous) lines.

Marker Assisted Selection and Marker Assisted Breeding

A breeding program can make use of marker assisted selection (MAS) andmarker assisted breeding (MAB) technologies to accelerate the successfuloutcome of a breeding project. These techniques enable theidentification of lines carrying a trait of interest in the laboratory,while other lines not containing a marker of interest can then bediscarded at an early stage. These methods can also increase theefficiency of a program, as the lines being evaluated in the field havea greater probability of meeting seed quality or other criteria. MAS andMAB methods rely on the existence of a dense set of genetic markers forthe species of interest. Genetic markers are the unique sequences thatmay be found in allelic forms of genes, distinguishing one allele fromanother. Like genes themselves, they can be transmitted to progeny in aMendelian fashion and can thus be used to follow the movement ofspecific alleles from parents to progeny.

Molecular markers can be used in Quantitative Trait Loci (QTL) mappingwhereby selection of plants with desired trait(s) is assisted by markersknown to be closely linked to alleles that have measurable effects on aquantitative trait—i.e., accumulation of markers linked to positiveeffecting alleles or elimination of markers linked to negative effectingalleles in the plant genome. Markers can also be used to select for thegenome of the recurrent parent and against the markers of the donorparent. This can minimize the amount of genome from a donor parent thatremains in the selected plants and/or the number of back crosses to therecurrent patent.

Other types of genetic markers in common use by persons skilled in theart include, but are not limited to, Restriction fragment lengthpolymorphisms (RFLP), Random Amplified Polymorphic DNA (RAPD), AmplifiedFragment Length Polymorphism (AFLP), Arbitrarily Primed Polymerase ChainReaction (AP-PCR), DNA Amplification Fingerprinting (DAF), SequenceCharacterized Amplified Regions (SCARs), Simple Sequence Repeats (SSRs),and Single Nucleotide Polymorphisms (SNPs).

Mutagenesis

Another method of creating genetic variation and capturing beneficialchanges in a heritable fashion is through mutagenesis breeding. Thismethod is often carried out via chemical means or ionizing radiation andis typically focused either on a microspore or on a whole seed level. InBrassica breeding, some common forms of mutagens used have been chemicalagents such as ethyl methanesulfonate (EMS) or N-ethyl-N-nitrosourea(ENU), or high levels of ionizing radiation (x-ray or gamma irradiationor exposure to UV light). EMS produces random point mutations via lowfrequency methylation of guanine residues in genomic DNA. This resultsin altered Watson-Crick base pairing such that the affected base pairingis converted from G-C to A-T. ENU is also an alkylating agent thatpreferentially modifies thymine residues converting A-T to G-C. Ionizingradiation may affect DNA in many ways but typically the mutations aredouble strand breaks leading to deletions and frameshift mutations thatare frequently inactivating.

To carry out this technique, seedlings or microspores are exposed to themutagenic agent and the surviving fraction are allowed to develop intomature plants. In some cases, the mutagenized plantlets or embryos (inthe case of microspore mutagenesis) may be exposed to selection in orderto enrich for a particular phenotype. For example, mutagenesis has beenused to develop plants that are resistant to the actions of specificherbicides; in this instance the developing plantlets or microspores canbe grown in vitro in the presence of the herbicide(s) of interest inorder to select for those plants with the appropriate mutationsconferring resistance. The advantage of the microspore mutagenesis ofthe seed approach is that the resultant DH plants can be used to derivepure and homozygous plant lines where all induced mutations, whetherdominant or recessive, would be expressed. Mutagenesis has been used todevelop Brassica varieties with resistance to various herbicides,altered seed oil profiles and increased tolerance to disease and abioticstress.

Genetic Transformation and Transgenic Technologies

In instances where unique and valuable traits are known to be availablein distant plant or in non-plant species that cannot be transferred toBrassica carinata via classical breeding, and where the genes for thosetraits have been cloned, a breeding program may employ genetictransformation techniques, or transgenic technologies, to stablytransfer those genes into this species. Transfer of cloned geneticelements into B. carinata have been achieved via a number of means,including PEG-mediated DNA uptake into protoplasts (Johnson, et al.,1989), electroporation into protoplasts (Fromm, et al., 1985), ballisticinfiltration using DNA coated microprojectiles (Finer, et al., 1999),Agrobacterium-based vector infiltration (Babic, et al., 1998), andinfection using plant virus-based vectors (Gleba, et al., 2004). Asidefrom having the genes of interest in cloned form, the other requirementsinclude having the genes cloned into a suitable vector to allow fortheir propagation in an appropriate bacterial system, as well as theirpackaging in appropriate viral and Agrobacterium strains iftransformation utilizes an infectious route of transfer. Oncetransferred, the gene(s) of interest would also require appropriateplant-based promoters, enhancers and terminators to allow for thecorrect temporal and tissue specific pattern of expression for theheterologous gene. Finally, to select for those rare events where theheterologous gene expression unit has been successfully transferred intothe plant genome, a selectable marker may be introduced, eitherphysically linked to the heterologous gene of interest or co-transformedwith the gene of interest at a suitable ratio to favor co-insertion.

The selectable marker may consist of a gene that can confer resistanceto a particular herbicide or antibiotic that would otherwise kill theplant, a gene that may confer a growth advantage, a gene that may altera response to plant hormones, or a gene that expresses a fluorescentprotein that can allow transformed cells to be easily visualized.Examples of selectable markers conferring resistance to antibiotics thathave been successfully used in Brassica transformation are the NPTIIgene (Bevan 1984; Datla, et al., 1992), encoding an enzyme conferringresistance to the antibiotic kanamycin, and the HPT gene encoding anenzyme conferring resistance to the antibiotic Hygromycin (Rothstein, etal., 1987). Examples of selection markers based on conferring toleranceto herbicides and successfully used in Brassica transformation are theBAR (Thompson, et al., 1987) and PAT (Wohlleben, et al., 1988) geneproducts, which encode phosphinothicine acetyltransferase and conferresistance to glufosinate (bialaphos) or L-PPT, and the AHAS geneproduct encoding acetohydroxyacid synthase enzyme conferring resistanceto imidazolinones (Miki, et al., 1990). Other plant selectable markershave been developed whose actions are not based on conferring resistanceto toxic compounds per se but instead allow survival in the presence ofnutrients not normally metabolized by the wildtype organism.

Transformation cannot only be used to introduce heterologous genes intothe genome of carinata plants, it can also be used to introduce nucleicacid constructs that are designed to modulate the expression ofendogenous genes. Nucleic acid constructs encoding antisense RNA or RNAisequences (Tang and Galili, 2004) can be used to interfere or knock downthe expression of endogenous genes to extremely low levels, simulatingthe effect of a null mutation at the endogenous locus. This of courserelies on the continuous stable expression of the antisense RNA or RNAito be effective. In amphidiploid Brassica species such as napus, junceaand carinata, multiple copies of genes from the contributing ancestralspecies may create a high level of functional redundancy such that asingle mutation in one of the homologues may not be sufficient to confera phenotype. However, by using an RNAi or antisense approach, where theinterfering RNA is derived from conserved sequences, one may conceivablybe capable of targeting all the expressed homologues and achieving afunctional knockdown effect.

Examples of modifications that can be introduced into Brassica carinatausing genetic transformation include, but are not limited to,

-   -   genes that control pollination, hybrid seed production, or        male-sterility;    -   genes encoding resistance to pathogens and insect pests (plant        disease resistance gene(s); gene(s) conferring resistance to        fungal pathogens; natural or synthetic Bacillus thuringiensis        (Bt) protein or a derivative thereof; an insect-specific hormone        or pheromone coach or a mimetic based thereon, or an antagonist        or agonist thereof; an insect-specific peptide that disrupts the        physiology of the affected pest; an enzyme responsible for        hyperaccumulation of a non-protein molecule with insecticidal        activity; a membrane permease, a channel former or a channel        blocker; a viral-invasive protein or a complex toxin derived        therefrom; an insect-specific antibody or an immunotoxin derived        therefrom; genes involved in the Systemic Acquired Resistance        (SAR) Response and/or the pathogenesis related genes; antifungal        genes; detoxification genes, such as for fumonisin, beauvericin,        moniliformin, and zearalenone, and their structurally related        derivatives; cystatin and cysteine protease inhibitors; Defensin        genes; genes that confer resistance to Phytophora Root Rot, such        as the Brassica equivalents of the Rps 1, Rps 1-a, Rps 1-b, Rps        1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2, Rps 3-a, Rps 3-b, Rps        3-c, Rps 4, Rps 5, Rps 6, Rps 7, and other Rps genes);    -   genes that confer resistance to a herbicide (mutant ALS and AHAS        enzymes to confer resistance to imidazolinone or sulfonylurea        herbicides; mutant 5-enolpyruvylshikimate-3-phosphate synthase        (EPSP) and aroA genes conferring resistance to glyphosate;        phosphinothricin acetyl transferase (PAT) and Streptomyces        hydroscopicus phosphinothricin-acetyl transferase (BAR)        conferring resistance to other phosphono compounds such as        glufosinate; ACCase inhibitor-encoding genes conferring        resistance to pyridinoxy or phenoxy propionic acids and        cyclohexones; pabA and gs+ genes conferring resistance to        triazine; nitrilase gene conferring resistance to benzonitrile;        acetohydroxy acid synthase, which has been found to make plants        resistant to multiple types of herbicide);    -   genes that confer or contribute to altered fatty acids        (down-regulation of stearoyl-ACP desaturase to increase stearic        acid; elevating oleic acid via FAD-2 gene modification and/or        decreasing linolenic acid via FAD-2 gene modification; altering        conjugated linolenic or linoleic acid content; altered LEC1,        AGP, Dek1, Superal1, mi1 ps, various 1pa genes such as 1pa 1,        lpa3, hpt, or hggt);    -   genes that confer or contribute to altered phosphorus content        (introduction of a phytase-encoding gene; up-regulation of a        gene that reduces phytate content);    -   genes that confer or contribute to altered carbohydrates,        antioxidant, or essential seed amino acids;    -   genes that create a site for site specific DNA integration, such        as the introduction of FRT sites that may be used in the FLP/PRT        system and/or Lox sites that may be used in the Cre/Loxp system;    -   genes that affect abiotic stress resistances (including, but not        limited to flowering, pod and seed development, enhancement of        nitrogen utilization efficiency, altered nitrogen        responsiveness, drought resistance or tolerance, cold resistance        or tolerance, and salt resistance or tolerance, and increase        yield under stress}.

Gene Editing

More recently, several novel approaches have been developed which offerthe ability to manipulate the plant genome in a targeted way.Collectively known as gene editing technology (Petolino, et al., 2010;Woo, et al., 2015, Sauer, et al., 2016), the technologies share severalimportant similarities:

-   -   they offer the ability to introduce small or large gene        deletions or insertions in the plant genome by means of targeted        double strand breaks at precise genomic locations;    -   they allow for the insertion of small inactivating mutations        into specific genes of interest;    -   they permit the replacement of an entire gene or gene segment by        a modified counterpart;    -   they allow for the insertion of a heterologous gene in a        specific genomic location which may represent a preferred site        for regulated gene expression. (i.e. downstream of a known        endogenous promoter/enhancer).

Locus Conversion

The Brassica carinata variety described herein represents a new basegenetic line into which a new locus or trait my be introduced usingtransgenic technologies or back-crossing. The term “locus conversion”refers to the product of such an introgression. For example, a donorparent having a specific desirable trait may be crossed with an inbredvariety, the recurrent parent, which has overall good agronomiccharacteristics yet lacks the desirable trait. The progeny of this crossis then mated back to the recurrent parent followed by selection of thedesired trait from the donor parent.

A locus converted plant cell of a locus converted plant is obtained byintroducing one or more locus conversion in the Brassica carinatavariety described herein. The locus converted plant cell is thereforeidentical to a cell from the Brassica carinata variety described hereinexcept for the one or more locus conversion and the locus convertedplant exhibits essentially all the physiological and morphologicalcharacteristics of the Brassica carinata variety described herein. Thelocus conversion can confer a trait selected from the group consistingof male sterility, disease resistance, fungal resistance, pestresistance, herbicide tolerance, abiotic stress tolerance, and alteredmetabolism. Both naturally occurring and transgenic DNA sequences may beintroduced through backcrossing. Molecular marker assisted breeding orselection maybe use used to reduce the number of backcrosses need toachieve the backcross conversion.

A locus conversion of the Brassica carinata variety described hereinwill otherwise retain its genetic integrity. For example, a locusconverted plant of the Brassica carinata variety described herein willcomprise at least 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of its basegenetics of the starting variety. A locus converted plant of theBrassica carinata variety described herein will therefore have at leastone, and possibly two or three physiological or morphologicalcharacteristics which are different from those of a locus convertedplant of the Brassica carinata variety described herein and otherwisehas all the physiological or morphological characteristics of a locusconverted plant of the Brassica carinata variety described herein.

Male Sterility

To produce and test hybrid combinations, some form of pollinationcontrol system must be employed. The most widely used pollinationcontrol system to achieve this in Brassica species is the method ofOgura (Ogura, 1968) adapted and modified by scientists at INRA, which isbased on control of pollination by use of male cytoplasmic sterility. Incytoplasmic male sterile (CMS) plants, a mitochondrial gene mutationinterferes with the flower's ability to produce viable pollen but doesnot affect the functionality of the flower's female components. Becausethe mutation is within the mitochondrial genome, it is transmittedmaternally through the cytoplasm. CMS plant lines, termed A lines, canonly produce viable seed via outcrossing with non-CMS plants; however,the F1 plants derived from the seed of such a crossing will also exhibitthe CMS phenotype and will still not produce viable seed by selfpollination.

The Ogura CMS trait, originally identified in Japanese radish varieties(Ogura, 1968), was transferred to Brassica oleracea via backcrossing(Bannerot, et al., 1977) then subsequently transferred to Brassicanapus. These initial Brassica napus A lines were found to be deficientin terms of their sensitivity to cold and their tendency to exhibit leafchlorosis, as well as fertility issues, which were ultimately found tobe related to interactions with chloroplasts derived from the radishcytoplasm (Pelletier et al., 1983). Using a cell fusion approach,improved Brassica napus A lines were subsequently developed thatovercame these Initial deficiencies (Pelletier, et al., 1987). A-linescan be maintained by crossing them with B lines (lines that aregenetically identical to A lines except that their cytoplasm is notCMS).

For production of fully fertile hybrid seed, CMS lines must be crossedwith lines that contain a restorer function gene (Rf) which can restorethe ability of CMS plants to produce viable pollen. Plant lines with theRF function stably incorporated and homozygous (termed RF lines) can becrossed with CMS lines and the F1 seed harvested from the CMS plantswill represent a true hybrid and which will have the ability to producefully fertile offspring capable of self pollination, a prerequisite forcommercial grain production.

Such an Rf activity, first identified in radish, was introduced intoBrassica napus (Heyn, 1976). It was subsequently shown that these lineswere affected by fertility problems (Heyn, 1978; Pellan-Delourme andRenard, 1988). Intensive backcrossing of these initial lines eventuallyyielded material with improved fertility such that they could be usedpractically as Rf lines (Delourme, et al., 1991). It was further noted,however, that hybrid seed produced from crosses with the early versionsof Brassica napus Rf lines often had high glucosinolate levels(Delourme, Foisset et al., 1998) that made generation of canola qualityhybrids problematic. Efforts to develop improved Rf lines by reducingthe amount of extraneous radish genetic material has resulted in newergenerations of Brassica napus (Primard-Brisset, et al., 2005) andBrassica juncea (Tian, et al., 2014) Rf line families of sufficientlyquality to be used in commercial hybrid seed production. Seed of Rflines can be maintained by allowing self crossing of plants grown inisolated tents or fields.

A, B and Rf lines are thus the prerequisite components of an expandableproduction system for F1 hybrid seed. In practice, for production ofpure hybrid seed:

-   -   a. the hybrid production fields are required to be        reproductively isolated from extraneous sources of Brassica        pollen;    -   b. RF lines are seeded at a predetermined optimal ratio relative        to A lines and may be kept spatially separated from the A lines        so that the RF plants can be readily removed post flowering, and        prior to harvest of the A line material;    -   c. pollinators are absolutely required to allow for maximal        pollination of the A lines by CMS pollen donors;    -   d. for maintenance and expansion of RF stocks, RF seed is        planted in isolation (either isolated fields separated from        other Brassica crops by great distance, or plots that are        reproductively isolated by virtue of a physical barrier such as        a tent) and allowed to self pollinate;    -   e. for maintenance and expansion of A line stocks, A lines and B        lines are planted in separate rows in isolated fields or tents        (as described above) and pollinators are used to promote        pollination of A lines with B line pollen. B lines are cut down        after flowering and before seed set and the A line plants are        allowed to come to maturity then harvested.

In some cases, a less labor- and cost-intensive alternative to thatdescribed in (b) is employed whereby a low ratio of RF plants may beinterspersed with the A line plants in the field and harvested together.While there will be some level of adulteration of the harvested hybridgrain with that of the self pollinated Rf plants; however, in practicethis level of contamination will not greatly affect the commercialquality of the hybrid seed

More recently, both the CMS and Rf traits have been transferred toBrassica carinata. The development of a set of genetically diverse CMSand Rf lines in Brassica carinata allows the testing of manycombinations for the first time in this Brassica species.

In a hypothetical test scheme based on the Ogura CMS system, thefollowing steps are carried out:

-   -   panels of genetically diverse Brassica carinata varieties are        selected from a previously characterized collection of elite        germplasm and used for introgression of CMS or Rf traits to        produce A and Rf test lines. Diversity can be estimated by        comparing differences in molecular marker profiles between        candidate lines;    -   crosses are carried out between selected diverse A and Rf test        lines, under conditions described in previous sections;    -   F1 carinata hybrid seed from test crosses are planted in        replicated small plot trials, F1 plants and harvested seed are        assessed for a number of traits including, but not limited to,        early vigour, yield potential, grain and oil yield per acre, as        well as seed quality traits;    -   patterns of heterosis between pairs A and Rf parents are        identified and F1 hybrid seed from promising combinations are        studied more extensively in larger, geographically diverse        trials; and    -   hybrid seed from specific combinations may be selected for        potential commercial release as a new variety.

Phenotype/Genotype

To be useful and reliable, a Brassica carinata variety or hybrid must behomogenous and reproducible. While there are a number of analyticalmethods known to the skilled person for assessing the phenotypicalstability of a Brassica carinata variety or hybrid, the traditionalmethod is the observation of phenotypic traits over the life of thecarinata plant using data collected from field experiments conductedunder the selected geographic, climatic, and soil conditions during oneor more growing seasons. Phenotypic characteristics observed include,but are not limited to, traits associated with seed yield (pod density,number of seeds per pod, pod length), oil yield, seed oil quality (GSLand erucic acid content; fatty acid composition), seed protein content,seed protein quality, glucosinolate composition of meal, growth habit,lodging resistance, plant height, and pod shatter resistance. Otherphenotypic characteristics that may be observed include, but are notlimited to, traits associated with pest tolerance or resistance, cold orfrost tolerance, disease tolerance or resistance, herbicide tolerance orresistance, early or late flowering, and/or early or late maturity.

In some embodiments, Brassica carinata varieties or hybrids useful forcommercial crop production or production of commercial products mayexhibit traits associated with seed yield, oil yield, seed oil quality,erucic acid content of seed or oil, glucosinolate content of seed, seedprotein content, fatty acid composition of oil, glucosinolatecomposition of meal, meal protein content, growth habit, lodgingresistance, plant height, and pod shatter resistance

In some embodiments, a Brassica carinata variety or hybrid may exhibitmultiple traits or phenotypic characteristics that provide agronomicadvantages in particular geographies, climates, cropping regimes, and/orsoil types. For example,

-   -   Brassica carinata varieties for planting in regions with a        climate classified as being of tropical moist characteristics,        with planting occurring in fall or winter for harvest in spring        or summer, may be selected for one or more traits including, but        not limited to, superior yield of oil per area planted, shorter        time to maturity, improved frost tolerance, improved disease        resistance, and resistance to pod shatter.    -   Brassica carinata varieties for planting in regions with a        climate classified as being of cool temperate, dry        classification, with planting occurring in spring and harvest in        summer or fall, may be selected for one or more traits        including, but not limited to, superior yield of oil per area        planted, shorter time to maturity, tolerance to drought,        improved disease resistance, and resistance to pod shatter.    -   Brassica carinata varieties for planting in regions with a        climate classified as being of warm temperate, moist        characteristic for planting in fall or winter for harvest in        spring or summer, may be selected for one or more traits        including, but not limited to, superior yield of oil per area        planted, shorter time to maturity, tolerance to drought,        improved disease resistance, and resistance to pod shatter.

Other traits for which Brassica carinata varieties may be selectedinclude, but are not limited to, Alternaria resistance, days toflowering, depth of canopy, duration of flowering, end of flowering,flower petal coloration, frost tolerance, herbicide resistance, leafcolour, leaf glaucosity, leaf length, leaf number of lobes, leaf width,maturity, seed maturity, number of seed-bearing pods, pod (silique) beaklength, pedicel length, petiole length, plant height, plant length, pod(silique) length, pod (silique) width, primary raceme length, recoveryfrom frost damage, Sclerotinia resistance, seed colour, seeds per pod,seed weight (thousand seed weight or TKW), and stem color.

Genotype assessment(s) can be used to confirm the homogeneity andreproducibility of a Brassica carinata hybrid, to identify plants of thesame variety or related variety, and to confirm the pedigree of theplant. Techniques known to those skilled in the art for the analysis andcomparison of plant genotype include, but are not limited to, wholegenome sequencing, Restriction fragment length polymorphisms (RFLP),Random Amplified Polymorphic DNA (RAPD), Amplified Fragment LengthPolymorphism (AFLP), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Simple Sequence Repeats (SSRs), and SingleNucleotide Polymorphisms (SNPs).

The Brassica carinata variety of the present invention has shownuniformity and stability for all traits described in the varietydescription information in Table 1, which include morphological,agronomic, and quality traits for Brassica carinata variety DH-040.008.The detailed phenotypic information provided in Table 1 is based on datacollected in field experiments using conventional agronomic practices.For comparative purposed, Brassica carinata varieties AAC-A120 andAGRO44-312D-HP11 (WO2017/181276A1; henceforth referred to as“AG044-HP11” or “HP11”) were similarly grown as comparative checkvarieties in replicated experiments, and observations were recorded forthe various morphological traits for Brassica carinata varietyDH-040.008 and the comparative check varieties.

Disease

Varieties of Brassica carinata may be susceptible to a number ofpathogens that commonly infect Brassica species including, but notlimited to, the bacteria and fungi listed below:

Disease Organism Disease Organism Bacterial leaf spot Pseudomonas Lightleaf spot Pyrenopeziza syringae brassicae Bacterial leaf rot Erwiniamarginalis Powdery mildew Erysiphe polygoni Bacterial soft rotPseudomonas Root rot complex Rhizoctonia solani, marginalis Fusariumspp., Pythium spp. Black rot Xanthomonas Seeding disease Rhizoctoniasolani, campestris complex Fusarium spp., Pythium spp. Alternaria blackspot Alternaria spp Sclerotonia white Sclerotonia stem rot sclerotiorumAlternaria grey leaf spot Alternaria brassicae Aster yellow Phytoplasmaspp. Alternaria brassicola Alternaria pos spot Alternaria brassicaeDamping off Phytophthora Alternaria brassicola cactorum Pythium spp.Black leg Leptosphaeria White leaf spot Mycosphaerella maculanscapsellae Leptosphaeria biglobosa Black root Aphanomyces Grey stemMycosphaerella raphanin capsellae Clubroot Plasmodiophora Wirestem andRhizoctonia solani brassicae girdling root rot Downey mildew PeronosporaWhite rust Albugo candida parasitica Fusarium wilt Fusarium oxysporumVerticillium wilt Verticillium Fusarium avenaceum longisporum

Examples of economically significant fungal diseases of Brassica speciesand mustard oilseeds include

-   -   a. Sclerotinia stem rot is caused by a fungus whose spores        infect Brassica species primarily during flowering stages and        whose incidence is associated with periods of high humidity.        Lesions are formed on the stems which can eventually kill the        plant. Fungicides are available which can control the severity        of the infection but must be applied at specific periods of the        plant lifecycle (i.e. at early to mid-flowering) for best        effect. Often multiple applications within this window of time        are necessary.    -   b. Alternaria is a fungal disease of Brassica species that        affects plants at all growth stages from early seedling through        to maturity although mature plants are more susceptible. The        greatest economic impact is on grain yield and quality. Foliar        fungicide application during the late flowering stage is an        effective way to mitigate the more detrimental effects of the        disease on grain yield and quality.    -   c. Blackleg is caused by a fungal pathogen Leptosphaeria        maculans of Brassica oilseed crops, which infects plants at all        stages, but early stage infections have the most serious        consequences, often culminating in plants with necrotic lesions        on their lower stems that can virtually sever the plants at the        base. Fungicides are only partially effective, having a minor        protective effect when applied at an early plant growth stage.    -   d. Clubroot is caused by a soil borne fungus-like pathogen        (Plasmodiophora brassicae) that affects the roots of Brassica        oilseed crops. The spores can persist for long periods in the        soil and there is currently no effective fungicidal treatment.        Management may require using rotations which limit the frequency        of Brassica planting.

The development of disease tolerant or disease resistant carinatavarieties is important for achieving high yields of seed, oil, and otherproducts from Brassica carinata. Conventional methods for control ofmicrobial disease may employ one or more of chemical control, diseaseresistance, and culture control procedures such as crop rotation,liming, and use of bait crops.

When producing a commercial crop or a group of cultivated Brassicacarinata plants in a field, harmful microorganisms can be controlled bythe application of a composition comprising one or more microbiocidalingredients, such as a fungicide. Fungicides comprise a diverse set ofchemical agents which can prevent or reduce the severity of plantinfection by pathogenic fungi. There are numerous classes of fungicides.FRAC (Fungicide Resistance Action Committee; http://www.frac.info/home)lists 12 classes based on the different biochemical pathways that thefungicides within a class targets, as well as a 13^(th) class whichcomprises fungicides with unknown modes of action. Fungicides are alsodistinguished by their modes of delivery and sites of action: somefungicides are sprayed onto plant surfaces, some are applied to the soilsurfaces either in granular form or as a liquid flooding the soilsurface, while others are applied as seed treatments. Fungicides appliedto the seeds or soils tend to be absorbed via the roots and aretransported to all plant tissues via xylem. Fungicides that are foliarcan be either local—i.e., protecting only the surfaces that theycontact, systemic—i.e., absorbed by the upper plant surfaces but thentransported by xylem to all above ground tissues, or partiallysystemic—i.e., they can be locally absorbed but can only be transportedshort distances to protect a somewhat more extensive surface than theinitial point of fungicide contact. Fungicides can help mitigate therisk of losses incurred by fungal infection, but the costs of fungicidespraying are significant enough to require cost benefit and riskassessment type analyses to be carried out before deciding to proceed.

When producing a commercial crop or a group of cultivated Brassicacarinata plants in a field, harmful microorganisms can be controlled bythe application of a composition comprising one or more microbicide orfungicide ingredient. Examples of microbiocides and fungicides usefulfor disease control for Brassica carinata include, but are not limitedto, azoxystrobin, boscalid, fluxapyroxad, pyraclostrobin, picoxystrobin,propiconazole, metconazole, iprodione, prothiaconazole, vinclozolin,carbathiin, thiram, difenoconazole, metalaxyl, sedaxane, fludioxonil,penflufen, trifloxystrobin, and sedaxane.

Insect Pests

A variety of insect pests may infest and/or cause damage to a developingBrassica carinata plant. Common insect pests of Brassica at the seedlingstage include, but are not limited to, aphids (Lipaphis erysimi, Myzuspersicae, Brevicoryne brassicae), flea beetles (Phyllotreta cruciferae,Phyllotreta striolata, Psylliodes punctulata), cutworm (Agrotisorthogonia, Euxoa ochrogaster, Feltia jaculifera, Lacinipolia renigrea),and cabbage root maggot (Delia radicum). Common insect pests of Brassicaat the flowering and podding stages include, but are not limited to,diamondback moth (Plutella xylostell), Berta armyworm (Mamestraconfigurata), and cabbage seed pod weevil (Ceutorhynchus obstrictus).

When producing a commercial crop or a group of cultivated Brassicacarinata plants in a field, insect pests may be controlled by theapplication of a composition comprising one or more insecticide.Insecticides are a group of pesticide compounds designed to reduce oreliminate crop loss due the predation of crop species by insects. Likeherbicides and fungicides, insecticides are classified according totheir mode of action and the biochemical pathways that they target. Oneclassification scheme (IRAC MoA) advocated by the Insecticide ResistanceAction Committee (IRAC; http://www.irac-online.org) lists 29 classes ofinsecticides grouped by the common biochemical processes and pathwaysthat the insecticide compounds target. Like herbicides and fungicides,insecticide function and persistence can also be influenced by theirsites of action, i.e. whether they are only active on the surface ofplants as applied, or whether they function as systemic agents. Furtherdifferentiation among some insecticide groups may be apparent based onwhether they exhibit selectivity for specific insect types due todistinctive aspects of that insect's biology. Given that some insectsserve a beneficial role, such as controlling plant pests, serving asplant pollinators and improving the nutrient content of soil, it isimportant that insecticides not be applied indiscriminately, but ratherare used in a way that limits their actions as much as possible to thedesired target species. Thus, modalities such as timing of application,amount and route of application, and restrictions on the types ofinsecticides used and the crops they may be used are all incorporatedinto the registered usage criteria of insecticide as a means of ensuringtheir safety and efficacy.

Insecticides useful for the control of insect pests of Brassica carinatainclude, but are not limited to, zeta-cypermethrim,zeta-cypermethrim-S-cyanol, lambdacyhalothrin, methoxyfenozide,cyantraniliprole, imidacloprid, thiamethoxam, and clothianidin. Suchinsecticide can be applied as a seed treatment or as a foliar treatment.

Plant Pests

Brassica carinata is an aggressive crop and will out-compete many weedsif it establishes well. Some weed species, however, if allowed toestablish early and persist, can affect quality and yield of all crops,including carinata. Examples of weeds that can adversely affect yieldand quality include cochia, wild mustard, and wild radish. Weedmanagement is thus an important aspect of modern agricultural practiceand comprises several different but complementary approaches includingphysical methods to remove weeds before seed can be set, such ascultivation, tilling and rogueing of fields as well as use of chemicalagents or herbicides to suppress or kill weedy species before theybecome established and/or are able to set and release their seed.

When producing a commercial crop or a group of cultivated Brassicacarinata plants in a field, plant pests can be controlled by theapplication of a composition comprising one or more herbicide.Herbicides comprise a large group of chemical compounds that interferewith specific biological processes of the plants in such a way as toblock their growth and survival. Herbicides are grouped into classesdefined by the biological process with which they interact. These caninclude inhibition of lipid biosynthesis, inhibition of amino acidbiosynthesis, hormonal regulation of plant growth, inhibition ofphotosynthesis, inhibition of nitrogen metabolism, inhibition of plantpigments biosynthesis or function, agents which can disrupt cellmembranes and agents which inhibit seedling growth (Sherwani, et al.,2015). In general, different compounds and herbicide classes may displaypreferential efficacy against certain weedy species. Moreover, some cropspecies may display more tolerance to certain classes of herbicide thanothers. Thus, in a particular geographical region, the use of aparticular herbicide for weed control may be dictated by the nature ofthe crop being cultivated and the native weeds encountered in theregion. The registered usage also specifies specific methods ofapplication of the herbicide, including recommended concentration ofherbicide, use of appropriate diluents, adjuvants, or surfactants,method of delivery (i.e. spray versus granular), timing of applicationat appropriate crop stage to ensure least crop damage, timing ofapplication and number of applications to ensure optimal weed control,location of application (foliar or soil application), recommendedweather conditions for optimal weed control. Some examples of herbicidesrecommended for use with Brassica carinata grown in SE USA are listed(Seepaul, et al., 2015).

Seed Cleaning

“Cleaning (of) seed” or “seed cleaning” refers to the removal of foreignmaterial from the surface of the seed. Foreign material to be removedincludes, but is not limited to, fungi, bacteria, insect material(including insect eggs, larvae, and parts thereof), and any other peststhat exist on the surface of the seed. “Cleaning (of) seed” or “seedcleaning” also refers to removal of any debris or low quality, infested,or infected seeds, or seeds of different species that are foreign to thesample.

Seed Treatment

Prior to planting, Brassica carinata a composition may be applied to theseed as a seed treatment. The composition may be applied at any timefrom harvesting of the seed to sowing of the seed using methodsincluding, but not limited to, mixing in a container, mechanicalapplication, tumbling, spraying, misting, and immersion. The compositionmay be applied as a liquid, a slurry, a mist, a soak, or a powder. Thecomposition may comprise one or more of a pesticide, fungicide,insecticide, antimicrobial, a bacterial or fungal inoculant for nutrientutilization, plant growth regulator, or plant signalling compound. Ageneral discussion of techniques for application of fungicides to seedsmay be found, for example, in chapter 9 of (Jeffs 1978)

Commercial Crops and Commercial Plant Products

Commercial crop production comprises the steps of seeding, cultivationand harvesting of grain:

-   -   Seeding: Brassica carinata can be planted into conventionally        tilled soil where conventional tillage or full-tillage comprises        a substantial soil inversion repeated several times yearly such        that few plant residues remain at the soil surface at the time        of seeding. Alternatively, carinata can planted into soil that        is maintained under conservation tillage practices whereby the        extent and frequency of tillage is substantially reduced with        respect to conventional tillage (so-called medium or low tillage        soil management) or preferably, it may be no-till planted in        standing stubble. Seeding is carried out at a rate designed to        achieve plant densities in a range from 80 to 180 plants per        square meter. B. carinata should be seeded at a consistent 1.25        to 2.5 cm depth. Brassica carinata is a mid- to long-season crop        that requires a slightly longer growing season than other        mustard types. Hence seeding early provides the best results.        The ideal seeding date depends greatly on geography and weather.        In general, soils should be at least 4° C. (40° F.) or higher        before planting. In the Canadian Prairies and US northern tier,        typical planting occurs in spring between early April to late        May. In South Eastern US, typical planting occurs in fall        between October and December. In South America, the optimal        planting time occurs in fall or winter (i.e. typically between        beginning of May to end of June).    -   Cultivation: For good stand establishment, Brassica carinata        requires adequate soil moisture at seeding and through emergence        but can tolerate reduced moisture thereafter and stands up well        to the semi-arid mid-summer conditions. Brassica carinata is a        temperate climate crop but which has been adapted to the more        extreme conditions experienced in the southern Canadian prairies        and Northern Tier US states. During initial stand formation,        carinata can recover from short term frost conditions and        tolerates higher heat during flowering and seed set better than        other Brassica oilseeds. The fertility requirements of Brassica        carinata are similar to other mustards and canola. Adequate        availability of the primary macronutrients nitrogen,        phosphorous, potassium and sulfur are required to achieve the        true yield potential. Lesser amounts of secondary        macronutrients, including calcium (Ca), magnesium (Mg) and        sulfur (S) and trace amounts of micronutrients (such as boron,        copper, Iron, manganese, zinc) may also contribute to optimal        plant growth and yield. Fertilizer rates vary with growing zone        and soil fertility.    -   Harvesting is the act of collecting the portion of a plant that        has matured sufficiently over the course of a growing season and        that has value as a source of food, feed, fibre, feedstock,        structural material or as a propagule for the plant itself.        Brassica carinata is harvested, for example, by mechanical        harvesting, ideally when seed maturity is reached (seed, pods        and stalks turn from green to yellow, seed moisture is 9.5 5 or        less). Brassica carinata can be combine harvested by straight        cutting or, if need be, can be swathed at an early stage,        allowed to dry naturally or with the aid of a desiccant, then        the dried swath can be combined. Swathing mean cutting near the        base of the plant and allowing the plant to lie flat in field        for several days to allow the grain to reach the appropriate        dryness. However, since Brassica carinata has a sturdy stalk,        the preferred method for harvest of carinata is direct combining        at maturity, rather than swathing or pushing followed by        combining.    -   Combining: refers to the process of reaping and collecting the        seed pods from the matured crop, threshing the seed pods to        release the seed (grain), and winnowing to separate and recover        the grain from the now empty seed pods, stems, and branches        (collectively referred to as chaff). These once distinct        operations are today often “combined” by use of a        multifunctional mechanized apparatus, appropriately known as a        “combine” harvester.    -   Grain, in reference to Brassica carinata, refers to the seed        harvested at maturity and sold as a source of oil and meal        products.

Commercial products produced from Brassica carinata seed include, butare not limited to, crushed, non-viable seed, oil, meal, and proteinisolate. Production of oil, meal and protein isolate involves multiplesteps. Typically, the seeds are cleaned then crushed in a roller mill togenerate flakes. The flaked seed then undergoes a cooking process inwhich it is conveyed to a heated drum where the flakes are cooked atelevated temperatures (typically from 70-90° C.). The cooking helps toreduce the viscosity of the oil to allow for more efficient extractionin subsequent steps. Cooked seed flakes are then pressed in a series ofscrew presses or expellers which can remove 50-60% of the oil. Asidefrom the oil, which is removed for further processing, the pressingproduces a meal cake that is ideal for solvent extraction. Using severalcycles of extraction, the meal cake is treated with a solvent such ashexane to remove the residual oil from the meal. The meal is thentransferred to a desolventizer-toaster where it is heated to removeremaining solvent; the final step of the process, called toasting,involves injection of stream into the meal to remove the last traces ofsolvent. The meal is then cooled and dried by blowing forced air throughit. In some cases, the seed can also be processed using a cold pressmethodology which is similar to above except it does not involve the useof solvent to remove residual oil from the oil cake, resulting in a mealwith much higher oil composition. In other cases, meal containing highlevels of glucosinolates can be combined with other ingredients andformulated into pellets for use as a biofumigant (U.S. Publication No.2008/0199451).

Brassica carinata plants can also be used commercially for biofumigationto reduce the population of disease-causing organisms in the soil.Typically, a field is seeded with Brassica plants early in the plantingseason, the plants are grown for a time, and the biomass is collectedbetween flowering and before seed set. The plant biomass is then choppedto release the myrosinase enzyme and convert the glucosinolates in theplant biomass to isothiocyanates. The chopped biomass is tilled into thesoil prior to seeding of a second crop

Uses of Brassica carinata Variety DH-040.008

Brassica carinata variety DH-040.008 can be used in accordance with anyof the breeding methods described herein, as well as in breeding methodsknown to those skilled in the art, to produce carinata hybrids or otherprogeny plants having the desired traits and characteristics of varietyDH-040.008.

The invention is directed to methods for producing carinata seeds,plants and plant parts from a carinata plant produced by crossing afirst parent plant with a second parent plant, wherein the first parentplant is Brassica carinata variety DH-040.008 and the second parentplant is also Brassica carinata variety DH-040.008, another Brassicacarinata variety, or a variety of another Brassicaceae species. In someembodiments, Brassica carinata variety DH-040.008 may be the male orfemale parent. In other embodiments, either the first or second parentplant may be male sterile. In some embodiments, the second parent plantcomprises a desired trait. In some embodiments, the desired trait isselected from the group consisting of male sterility, diseaseresistance, fungal resistance, pest resistance, herbicide tolerance,abiotic stress tolerance, and altered metabolism. In other embodiments,the desired trait is herbicide tolerance and the herbicide is selectedfrom, but not limited to, the group consisting of glyphosate,glufosinate, imidazolinones, and auxin analogues such as 2,4-D anddicamba.

In another aspect, the invention is directed to a method of producing afirst generation (F1) hybrid Brassica carinata seed, as well a firstgeneration (F1) hybrid plant grown from such seed, comprising crossingBrassica carinata variety DH-040.008 with a different Brassica carinataplant and harvesting the resultant F1 hybrid Brassica carinata seed, andwherein Brassica carinata variety DH-040.008 is either a female parentor a male parent.

In another aspect, the invention is further directed to a method forproducing a Doubled Haploid (DH) variety comprising isolating a flowerbud of the F1 hybrid plant, dissecting out a haploid microspore, placingthe haploid microspore in culture, inducing the microspore todifferentiate into an embryo and subsequently into a plantlet,identifying whether the plantlet contains a diploid chromosome number,wherein the diploid chromosome number occurred through chromosomedoubling, and continuing to grow the plantlet if it contains a diploidchromosome number. In one embodiment, the diploid chromosome number isobtained by chemical or physical means. In another aspect the inventionis directed to a plant, plant part, or seed of such Double Haploidvariety produced from the F1 hybrid plant resulting from crossingBrassica carinata variety DH-040.008 with a different Brassica carinataplant.

In another aspect, the invention is directed to producing a Brassicacarinata plant, as well as a plant, plant part, cell, or seed therefrom,by crossing Brassica carinata variety DH-040.008 with a second Brassicacarinata variety having a desired trait, growing the resultant F1 hybridseed and selecting one or more progeny plants that have the desiredtrait, backcrossing the selected progeny plants that have the desiredtrait with plants of variety DH-040.008 to produce backcross progenyseed, and growing the resultant backcross progeny seed and selectingbackcross progeny plants that have the desired trait and at least aportion of the genetic make up of Brassica carinata variety DH-040.008.nIn some embodiments, the steps of backcrossing and growing the resultingprogeny seed may be repeated 1 to 2 times, 1 to 3 times, 1 to 4 times,or 1 to 5 times, or until the Brassica carinata variety produced fromvariety DH-040.008 has the desired trait and the physiological and/ormorphological characteristics set forth in one or more of Tables 1, 2,3, 4, and 5, as determined at the 5% significance level, when grown inthe same location under the same environmental conditions as varietyDH-040.008. In some embodiments, the desired trait is selected from thegroup consisting of male sterility, disease resistance, fungalresistance, pest resistance, herbicide tolerance, abiotic stresstolerance, and altered metabolism. In other embodiments, the desiredtrait is herbicide tolerance and the herbicide is selected from, but notlimited to, the group consisting of glyphosate, glufosinate,imidazolinones, and auxin analogues such as 2,4-D and dicamba.

In yet another aspect, the invention is directed to a method ofproducing a Brassica carinata variety having a desired trait, as well asseeds, plants and plant parts of such variety, by crossing a plant ofBrassica carinata variety DH-040.008 with a plant of another species ofthe family Brassicaceae comprising the desired trait, producing F1progeny plants using embryo rescue techniques to recover viable F1plants or growing F1 seeds, self-pollinating the F1 plants that have thedesired trait and carinata character, producing F2 plants using embryorescue techniques to recover viable F2 plants or growing F2 seeds,self-pollinating the F2 plants that have the desired trait and carinatacharacter, using embryo rescue techniques to recover viable F3 plants orgrowing F3 seeds to produce progeny plants, self-pollinating the progenyplants that have the desired trait and carinata character to producefurther progeny plants, and selecting the progeny plants with thedesired trait and carinata character. In some embodiments, the steps ofproducing progeny plants, self-pollinating, and selecting progeny plantshaving the desired trait and carinata character are repeated until theprogeny plant has the desired trait and the physiological and/ormorphological characteristics set forth in one or more of Tables 1, 2,3, 4, and 5, as determined at the 5% significance level, when grown inthe same location under the same environmental conditions as varietyDH-040.008. In other embodiments, the steps of backcrossing and growingthe resulting progeny seed may be repeated 1 to 2 times, 1 to 3 times, 1to 4 times, or 1 to 5 times, or until the Brassica carinata varietyproduced from Brassica carinata variety DH-040.008 has the desired traitand the physiological and/or morphological characteristics set forth inone or more of Tables 1, 2, 3, 4, and 5, as determined at the 5%significance level, when grown in the same location under the sameenvironmental conditions as variety DH-040.008. In some embodiments, thedesired trait is selected from the group consisting of male sterility,disease resistance, fungal resistance, pest resistance, herbicidetolerance, abiotic stress tolerance, and altered metabolism. In otherembodiments, the desired trait is herbicide tolerance and the herbicideis selected from, but not limited to, the group consisting ofglyphosate, glufosinate, imidazolinones, and auxin analogues such as2,4-D and dicamba.

In another aspect, the present invention is directed to a cell of aplant of a Brassica carinata variety having a desired trait, as well asseeds, plants and plant parts of such variety, by crossing a plant ofBrassica carinata DH-040.008 with a plant of another species of thefamily Brassicaceae comprising the desired trait, producing F1 progenyplants using embryo rescue techniques to recover viable F1 plants orgrowing F1 seeds, self-pollinating the F1 plants that have the desiredtrait and carinata character, producing F2 plants using embryo rescuetechniques to recover viable F2 plants or growing F2 seeds,self-pollinating the F2 plants that have the desired trait and carinatacharacter, using embryo rescue techniques to recover viable F3 plants orgrowing F3 seeds to produce progeny plants, self-pollinating the progenyplants that have the desired trait and carinata character to producefurther progeny plants, and selecting the progeny plants with thedesired trait and carinata character. In some embodiments, the steps ofproducing progeny plants, self-pollinating, and selecting progeny plantshaving the desired trait and carinata character are repeated until theprogeny plant has the desired trait and the physiological and/ormorphological characteristics set forth in one or more of Tables 1, 2,3, 4, and 5, as determined at the 5% significance level, when grown inthe same location under the same environmental conditions as varietyDH-040.008. In some embodiments, the desired trait is selected from thegroup consisting of male sterility, disease resistance, fungalresistance, pest resistance, herbicide tolerance, abiotic stresstolerance, and altered metabolism. In other embodiments, the desiredtrait is herbicide tolerance and the herbicide is selected from, but notlimited to, the group consisting of glyphosate, glufosinate,imidazolinones, and auxin analogues such as 2,4-D and dicamba.

Another aspect of the present invention is directed to a tissue cultureof protoplasts or regenerable cells of the plant or plant part producedfrom the seed of Brassica carinata variety DH-040.008, as well as aBrassica carinata plant regenerated from the tissue culture thephysiological and/or morphological characteristics set forth in one ormore of Tables 1, 2, 3, 4, and 5, as determined at the 5% significancelevel, when grown in the same location under the same environmentalconditions as variety DH-040.008. The tissue may be selected from thegroup consisting of leaves, pollen, embryos, roots, root tips, pods,flowers, ovules, and stalks.

In other aspects, the present invention is directed to a cell ofBrassica carinata variety designated DH-040.008, as well as to a plantor plant part, or a protoplast of a plant or plant part, produced from aseed of Brassica carinata variety designated DH-040.008. In someembodiments, the plant part is an ovule, a leaf, pollen, a seed, anembryo a root, a root tip, a pod, a flower, or a stalk. In otherembodiments, the plant part is pollen or an ovule. In other embodiments,the present invention is directed to a cell, plant or plant part havingthe physiological and/or morphological characteristics set forth in oneor more of Tables 1, 2, 3, 4, and 5, as determined at the 5%significance level, when grown in the same location under the sameenvironmental conditions as variety DH-040.008.

The present invention also includes a method of producing a Brassicacarinata variety comprising a desired trait, as well as a plant, plantpart, or seed of such variety, by introducing a nucleic acid constructconferring the desired trait into a Brassica carinata plant of varietyDH-040.008 using polyethylene glycol (PEG) mediated uptake,electroporation, ballistic infiltration using DNA coatedmicroprojectiles (gene gun), an Agrobacterium infiltration-based vector,or a plant virus-based vector. In some embodiments, the nucleic acidconstruct comprises a transgene. In other embodiments, the nucleic acidconstruct comprises an RNAi construct. In some embodiments, the Brassicacarinata variety comprises the desired trait and the physiologicaland/or morphological characteristics set forth in one or more of Tables1, 2, 3, 4, and 5, as determined at the 5% significance level, whengrown in the same location under the same environmental conditions asvariety DH-040.008. In other embodiments, the desired trait is selectedfrom the group consisting of male sterility, disease resistance, fungalresistance, pest resistance, herbicide tolerance, abiotic stresstolerance, and altered metabolism. In other embodiments, the desiredtrait is herbicide tolerance and the herbicide is selected from, but notlimited to, the group consisting of glyphosate, glufosinate,imidazolinones, and auxin analogues such as 2,4-D and dicamba.

In another aspect, the invention is directed to the use of a plant ofBrassica carinata variety DH-040.008 to produce a Brassica carinatavariety comprising a new trait, wherein the new trait is introduced byexposing seedlings or microspores to a mutagenic agent. In someembodiments, the mutagenic agent is ethyl methanesulfonate,N-ethyl-N-nitrosourea, or x-ray, gamma or ultraviolet radiation.

In another aspect, the invention is directed to a cell of a plant of aBrassica carinata variety produced from variety DH-040.008, wherein theBrassica carinata variety comprises a new trait and is produced by amethod comprising exposing seedlings or microspores to a mutagenic agentand allowing the surviving fraction to develop into mature plants. Insome embodiments, the mutagenic agent is ethyl methanesulfonate,N-ethyl-N-nitrosourea, or x-ray, gamma or ultraviolet radiation.

In another aspect, the invention is directed to a method of producing acommercial crop of a Brassica carinata variety DH-040.008 or of aBrassica carinata variety produced by any of methods described herein.In another aspect, the invention is directed to a method of producing acommercial plant product from the commercial crop. In some embodiments,the commercial plant product comprises oil, meal, protein isolate, orbiofumigant. In another aspect the invention is directed to a method ofproducing crushed, non-viable seed of Brassica carinata varietyDH-040.008 or a Brassica carinata variety produced by any of methodsdescribed herein.

In another aspect, the invention is directed to a method ofbiofumigation comprising, growing a plant of Brassica carinata varietyDH-040.008 or a Brassica carinata variety produced by any of methodsdescribed herein in a field, collecting the Brassica carinata plantbiomass between flowering and seed set, chopping the biomass, andincorporating the chopped biomass into the soil.

The citation of any publication herein is not an admission that thepublication is prior art with respect to the present application.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it is readily apparent to those of ordinary skill in theart in light of the teachings of this invention that certain changes andmodifications may be made thereto without departing from the scope ofthe appended claims.

It is to be understood that any numerical value inherently containscertain errors necessarily resulting from the standard deviation foundin the respective testing measurements. Also, as used herein, the term“about” generally means within 10%, 5%, 1%, or 0.5% of a given value orrange. Alternatively, the term “about” means within an acceptablestandard error of the mean when considered by one of ordinary skill inthe art. Unless indicated to the contrary, the numerical parameters setforth in the present disclosure and attached claims are approximationsthat can vary as desired. At the very least, each numerical parametershould at least be construed in light of the number of reportedsignificant digits and by applying ordinary rounding techniques.

As used in this specification and the appended claims, the singularforms “a,” “an,” and “the” include plural reference unless the contextclearly dictates otherwise. Unless defined otherwise all technical andscientific terms used herein have the same meaning as commonlyunderstood to one of ordinary skill in the art to which this inventionbelongs.

The phrase “and/or,” as used herein in the specification and in theclaims, should be understood to mean “either or both” of the elements soconjoined, i.e., elements that are conjunctively present in some casesand disjunctively present in other cases. Multiple elements listed with“and/or” should be construed in the same fashion, i.e., “one or more” ofthe elements so conjoined. Other elements may optionally be presentother than the elements specifically identified by the “and/or” clause,whether related or unrelated to those elements specifically identified.Thus, as a non-limiting example, a reference to “A and/or B”, when usedin conjunction with open-ended language such as “comprising” can refer,in one embodiment, to A only (optionally including elements other thanB); in another embodiment, to B only (optionally including elementsother than A); in yet another embodiment, to both A and B (optionallyincluding other elements); etc.

As used herein in the specification and in the claims, “or” should beunderstood to encompass the same meaning as “and/or” as defined above.For example, when separating items in a list, “or” or “and/or” shall beinterpreted as being inclusive, i.e., the inclusion of at least one, butalso including more than one, of a number or list of elements, and,optionally, additional unlisted items.

As used herein, whether in the specification or the appended claims, thetransitional terms “comprising”, “including”, “carrying”, “having”,“containing”, “involving”, and the like are to be understood as beinginclusive or open-ended (i.e., to mean including but not limited to),and they do not exclude unrecited elements, materials or method steps.Only the transitional phrases “consisting of” and “consistingessentially of”, respectively, are closed or semi-closed transitionalphrases with respect to claims and exemplary embodiment paragraphsherein. The transitional phrase “consisting of” excludes any element,step, or ingredient which is not specifically recited. The transitionalphrase “consisting essentially of” limits the scope to the specifiedelements, materials or steps and to those that do not materially affectthe basic characteristic(s) of the invention disclosed and/or claimedherein.

Examples Example 1: Field Trials

Small plot field experiments were conducted in Canada and northernstates of the United States and Canada using best agronomic practicesfor carinata as described in the “Carinata Management Handbook” for theNorthern Plains(http://growcarinata.com/ckfinder/userfiles/files/US%20Northern%20Plains_handbook_2018.pdf).Several newly developed carinata varieties, including DH-040.008, weregrown in a Randomized Complete Block Design trial (RCBD), comprising 4replicate plots (unless otherwise noted) along with plots of checkvarieties AAC-A120 and/or AGRO44-312D-HP11 (henceforth referred to as“AGRO44-HP11” or “HP11”). During the trials, the experimental lines andcheck varieties were closely observed, and descriptions of phenotypiccharacteristics and agronomic traits were recorded. After plants hadreached maturity, seed was collected for subsequent NIR based seedquality analysis (as described in the definitions section).

Example 2: Varietal Characteristics of DH-040.008 Grown in Northern USand Canada

The northern tier states of the US, such as North Dakota and itsneighbours, as well as the adjacent southern Canadian prairie regions ofAlberta and Saskatchewan, constitute an important Brassica oilseedgrowing region. Evaluation of new carinata varieties in this regionallows for identification of varieties best adapted to growing in thelocal environment, comprising a semiarid environment with warm summerand cold winter. Under such conditions, carinata varieties productiveunder long daylight spring-summer cultivation in semiarid environmentswould prove advantageous to local farmers. Field trials are carried outannually at several sites in the region as described in Example 1 toidentify best candidates for commercialization based on agronomictraits, seed yield and quality as well as multi-year performance.

The detailed phenotypic information provided in Table 1 is based on datacollected from small plot, RCBD field experiments conducted in northernUS states and Canada, as described in Example 1. Each characteristic wasassessed as described in DEFINITIONS section, above. Least square meanvalues from replicated data sets were calculated using the REML modeland pairwise comparisons carried out using Tukey's test at the 5%probability level. All quantitative data analysis was carried out usingIMP 13 (SAS). Paired LSM values that share a common letter (lettercolumns of Table 1) are not significantly different from one another.

TABLE 1 Agronomic characteristics of DH-040.008 in trials in northern USand Canada Botanical name Brassica carinata Season type Spring/winterPollination control Open pollinated Year 2017 2018 DH- AAC- DH- AAC-Variety 040.008 Letters A120 Letters 040.008 Letters A120 Letters Daysto first flower 51.5 AB 48.6 CD 57.5 ABC 56.0 ABC Days to last flower73.0 ND 77.3 ND 75.5 ABC 73.0 ABC Duration of flowering 24.0 ND 31.3 ND18.0 A 17.0 A (days) Days to maturity 107.1 A 106.4 A ND ND Top ofcanopy (cm) 102.2 AB 96.7 AB 78.00 A 84.25 A Bottom of canopy (cm) 48.6AB 46.4 AB 28.25 AB 33.25 AB Depth of canopy (cm) 56.5 AB 53.1 AB 49.75A 51.00 A Lodging resistance 5.9 ND 5.1 ND 7.0 A 7.0 A (1-7) Pod shatter4.3 ND 4.8 ND 4.0 AB 3.5 AB resistance (1-7) ND = not determined;

Relative to check variety AAC-A120, Brassica carinata variety DH-040.008shows the following agronomic characteristics:

Flowering In the 2017 trials, plants of DH-040.008 initiated floweringsignificantly later than the check variety but had a shorter floweringperiod. In the 2018 trials, plants of DH-040.008 initiated floweringonly slightly later than the check variety but had a similar floweringperiod Maturity In the 2017 trials, plants of DH-040.008 reachedmaturity at the same time as the check variety. Canopy In both the 2017and 2018 trials, plants of DH-040.008 were slightly taller in staturethan check variety. The bottom of canopy of DH-040.008 plants was eithera bit higher (2017) or a bit lower (2018) than for plants of the checkvariety. However, depth of canopy slightly larger for DH-040.008relative to check variety AGR044-HP11 in both 2017 and 2018 trials.Lodging Under the growth conditions of the trial, plants of DH-040.008exhibited similar or slightly greater resistance to lodging compared toplants of the check variety. Pod shatter Pods from plants of DH-040.008exhibited either a slightly lower resistance (2017) or slightly higher(2018) resistance to shattering compared to pods from plants of thecheck variety.

Agronomic data accumulated from the trials described above indicate thatthe DH-040.008 variety possesses traits consistent with those of anexisting commercial line typically grown in the northern US and Canadiansouthern prairies and supports selection of DH-040.008 as suitablecandidate for commercial cultivation in this region.

Example 3: Seed Yields of Variety DH-040.008 from Trials in Northern USand Canada

Tables 2 and 3 (below) summarize, respectively, the results of yieldanalysis carried out on DH-040.008 and check variety AAC-A120 in 2016,2017, and 2018. Least square means (LSM) were calculated from replicatetrials for each variety tested at each site using a restricted maximallikelihood (REML) estimate, which can better describe data sets wheresome replicate data might be missing due to weather or disease relatedlosses. Tukey's test was then applied pairwise to determine whetherthere were statistically significant differences between the LSM yieldsfor DH-040.008 and the check variety. An aggregate of the yield datafrom all sites was obtained and the aggregated LSM yield values for eachvariety were compared to one another as described above. Paired LSMvalues that share a common letter (letter columns of Tables 2 and 3) arenot significantly different from one another.

TABLE 2 Seed yields of DH-040.008 from 2016 trials in northern US statesand Canada Site Variety LSM Yield (kg/ha) Letters Medicine Hat, ABDH-040.008 1551 D AAC-A120 1805 ABCD Devils Lake, ND DH-040.008 4190 ABAAC-A120 3130 ABCD Moosomin, SK DH-040.008 3330 AB AAC-A120 2705 ABCDEAll sites DH-040.008 3004 A AAC-A120 2547 AB

TABLE 3 Seed yields of DH-040.008 from 2017 trials in northern US statesand Canada Site Variety LSM Yield (kg/ha) Letters Carrington, NDDH-040.008 2504 ABC AAC-A120 2086 BC Indian Head, SK DH-040.008 3318 AAAC-A120 3057 A Milestone, SK DH-040.008 1744 AB AAC-A120 1827 ABOutlook, SK DH-040.008 3471 A AAC-A120 3434 A All sites DH-040.008 2773A AAC-A120 2601 AB

While the yield data reflect considerable variation between sites, ateach individual site the difference in LSM yields between DH-040.008 andcheck variety AAC-A120 were small and found not to be significantlydifferent.

The data from trials carried out in northern US states and Canadiansouthern prairies in 2016 (Table 2) and 2017 (Table 3) demonstratesimilar yield potentials for DH-040.008 and a check variety AAC-A120,supporting selection of DH-040.008 as a candidate for commercialcultivation in this region.

Example 4: Quality of carinata seed harvested from field trialsconducted in northern US and Canada

Trials were conducted as described in Example 1. Seed quality data wereobtained from NIR analysis as described above under DEFINITIONS, onreplicated samples of harvested seed from small plot, RCBD fieldexperiments. Least square means (LSM) were calculated for measurementsof replicate samples using a restricted maximal likelihood (REML)estimate, which can better describe data sets where some replicate datamight be missing due to weather or disease related losses. Tukey's testwas then applied pairwise to determine whether there were statisticallysignificant differences between the LSM yields for DH-040.008 and thecheck variety (Table 4). Paired LSM values that share a common letter(letter columns of Table 4) are not significantly different from oneanother.

TABLE 4 Seed quality characteristics of DH-040.008 harvested from fieldtrials in northern US and Canada. 2016 2017 2018 Quality Variety LSMLetters LSM Letters LSM Letters Seed Protein content, DH-040.008 28.31ABC 28.51 CD 33.86 A wt % of seed AAC-A120 28.48 ABC 29.35 BC 34.52 ASeed Oil content, DH-040.008 44.61 ABB 43.70 BC 40.13 ABC wt % of seedAAC-A120 44.29 ABC 43.12 BC 39.34 ABC Glucosinolates, DH-040.008 78.44CD 89.43 D 97.59 BCDEFG μmol/g AAC-A120 80.62 ABCD 102.9 B 110.40 ABCDEErucic Acid, DH-040.008 41.13 ABC 43.45 BC ND wt % of fatty acidsAAC-A120 40.46 ABCDEFG 43.04 CD ND Total Saturates, DH-040.008 5.73 IJ5.73 EF ND wt % of fatty acids AAC-A120 5.88 DEFGHIJ 5.71 EF ND ND = notdetermined

The fatty acid profile of seed oil from DH-040.008, as determined by NIRanalysis, was also compared to that of the AAC-A120 check variety andthe results, expressed as the percentage by weight (mean of 4 samples)for each of the major fatty acid constituents of the oil are shown inTable 5. Paired LSM values that share a common letter (letters column ofTable 5) are not significantly different from one another.

TABLE 5 Fatty acid profile of variety DH-040.008 grown in Canada in 2018Fatty Acid Variety LSM Letters C18:1 DH-040.008 4.02 DEFG AAC-A120 4.60CDEF C18:2 DH-040.008 14.00 DEF AAC-A120 14.93 BCDEF C18:3 DH-040.00815.18 BCDE AAC-A120 15.37 ABCDE C20:1 DH-040.008 7.13 ABCDE AAC-A1207.71 ABCDE

Seed oil content In the trials described above, seed from DH-040.008has, on average, statistically similar oil content as seed from checkvariety AAC-A120. Seed protein content In the trials described above,seed from DH-040.008 has, on average, statistically similar proteincontent as seed from check variety AAC-A120. Glucosinolates In thetrials described above, seed from DH-040.008 has, on average,statistically similar or somewhat higher glucosinolate content than seedfrom check variety AAC-A120. Erucic acid In the trials described above,oil from DH-040.008 seed has statistically similar levels of erucic acidcompared to oil from seed of check variety AAC-A120. Fatty acid profileIn the trials described above, oil from DH-040.008 seed has astatistically similar fatty acid profile as oil from seed of checkvariety AAC-A120.

As can be seen in Tables 4 and 5, seed from DH-040.008 showed a similaroil content as that of the commercial check variety which, based onyield equivalence between DH-040.008 and the check variety (shown inExample 3), could result in similar amounts of oil recovered per hectarefor fields planted with DH-040.008 and current commercial varieties.

DH-040.008 seed produced in this trial contained a similar proteincontent as that of the check variety. Meal produced from processing ofthis seed would therefore be expected to contain >40 wt % proteincontent, representing an advantage with respect to competing Brassicaoil seed meals such as canola meal.

Based on the potential for oil yields equivalent to oil yields from theexisting commercial variety, as well as the seed quality attributesdiscussed above, the DH-040.008 variety may be a suitable candidate forcommercial exploitation in the northern US states and Canada, andpossibly other geographical regions.

DEPOSIT

Applicant(s) have made a deposit of at least 2500 seeds of Brassicacarinata variety DH-040.008 with the NCIMB Ltd., Ferguson Building,Craibstone Estate, Bucksburn, Aberdeen, UK, AB21 9YA, under Accessionnumber 42999. The seeds deposited with NCIMB on Apr. 3, 2018 forDH-040.008 were taken from the seed stock maintained by AgrisomaBiosciences Inc., since prior to the filing date of this application.The deposit of seed of Brassica carinata variety DH-040.008 will bemaintained in the NCIMB depository, which is a public depository, for aperiod of 30 years, or 5 years after the most recent request, or for theenforceable life of the patent, whichever is longer, and will bereplaced if it becomes nonviable during that period. Additionally,Applicant has satisfied all the requirements 37 C.F.R. § 1.801-1.809,including providing an indication of the viability of the sample upondeposit. Applicant imposes no restrictions on the availability of thedeposited material from NCIMB; however, Applicant has no authority towaive any restrictions imposed by law on the transfer of biologicalmaterial or its transportation in commerce. Applicant(s) do not waiveany infringement of their rights granted under this patent or rightsapplicable to Brassica carinata variety DH-040.008 under the PlantVariety Protection Act (7 USC 2321 et seq.).

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1. A Brassica carinata variety DH-040.008, representative seed of saidvariety having been deposited under NCIMB accession number
 42999. 2. Aseed, plant, plant part or cell of Brassica carinata variety DH-040.008,representative seed of said variety having been deposited under NCIMBaccession number
 42999. 3. A Brassica carinata plant or plant parthaving the physiological and/or morphological characteristics set forthin one or more of Tables 1, 2, 3, 4, and 5, as determined at the 5%significance level, when grown in the same location under the sameenvironmental conditions as variety DH-040.008.
 4. A Brassica carinataseed produced by crossing Brassica carinata plants and harvesting theresulting Brassica carinata seed, wherein at least one Brassica carinataplant is the plant of claim
 2. 5. A method of producing a Brassicacarinata variety derived from Brassica carinata variety DH-040.008, themethod comprising (c) crossing a plant of Brassica carinata varietyDH-040.008 with a different Brassica carinata plant having a desiredtrait to produce F1 hybrid seed; and (d) growing the resultant F1 hybridseed and selecting one or more F1 hybrid plants having the desired traitand at least a portion of the genetic make up of Brassica carinatavariety DH-040.008.
 6. The method of claim 5, further comprising thesteps of (a) backcrossing the selected F1 hybrid plants with plants ofvariety DH-040.008, or with the different Brassica carinata plant havinga desired trait, to produce backcross progeny seed; (b) growing theresultant backcross progeny seed and selecting backcross progeny plantsthat have the desired trait and at least a portion of the genetic makeup of Brassica carinata variety DH-040.008; and (c) repeating steps (a)and (b) on the selected backcross progeny plants to a maximum of 10generations to produce a progeny Brassica carinata plant derived fromBrassica carinata variety DH-040.008, wherein the progeny Brassicacarinata plant comprises the desired trait and at least a portion of thegenetic make up of Brassica carinata variety DH-040.008.
 7. The methodof claim 5, further comprising the steps of (a) self-pollinating theselected F1 hybrid plants to produce further progeny seed; (b) growingthe further progeny seed and selecting further progeny plants that havethe desired trait and at least a portion of the genetic make up ofBrassica carinata variety DH-040.008; and (c) repeating steps (a) and(b) on the selected further progeny plants to a maximum of 10generations to produce a progeny Brassica carinata plant derived fromBrassica carinata variety DH-040.008, wherein the progeny Brassicacarinata plant comprises the desired trait and at least a portion of thegenetic makeup of Brassica carinata variety DH-040.008.
 8. An F1 hybridplant grown from the F1 hybrid seed produced by the method of claim 5,wherein the F1 hybrid plant has a desired trait and at least a portionof the genetic make up of Brassica carinata variety DH-040.008.
 9. Amethod for producing a Double Haploid variety comprising: (a) isolatinga flower bud of the F1 plant of claim 8; (b) dissecting out a haploidmicrospore; (c) placing the haploid microspore in culture; (d) inducingthe microspore to differentiate into an embryo and subsequently into aplantlet; (e) identifying whether the plantlet contains a diploidchromosome number, wherein the diploid chromosome number occurredthrough chromosome doubling; and (f) continuing to grow the plantlet ifit contains a diploid chromosome number, wherein the Double Haploidvariety comprises at least a portion of the genetic makeup of Brassicacarinata variety DH-040.008.
 10. A cell, seed, plant, or plant part ofthe Double Haploid variety produced by the method of claim
 9. 11. A cellof a Brassica carinata plant or plant part of claim
 3. 12. A tissueculture of protoplasts or regenerable cells of the cell of claim
 11. 13.The tissue culture of protoplast or regenerable cells of claim 12,wherein the protoplasts or regenerable cells are produced from a tissueselected from the group consisting of leaves, pollen, embryos, roots,root tips, pods, flowers, ovules, and stalks.
 14. A Brassica carinataplant regenerated from the tissue culture of claim 12, wherein the planthas the physiological and/or morphological characteristics set forth inone or more of Tables 1, 2, 3, 4, and 5, as determined at the 5%significance level, when grown in the same location under the sameenvironmental conditions as variety DH-040.008.
 15. A method ofproducing a commercial plant product comprising growing the plant ofclaim 2 to produce a commercial crop and producing the commercial plantproduct from the commercial crop.
 16. A method of producing a commercialplant product comprising growing the F1 hybrid plant of claim 8 toproduce a commercial crop and producing the commercial plant productfrom the commercial crop.
 17. A method of producing a commercial plantproduct comprising growing a plant of the Double Haploid variety ofclaim 10 to produce a commercial crop and producing the commercial plantproduct from the commercial crop.
 18. A commercial plant productproduced from a Brassica carinata plant of variety DH-040.008, whereinthe commercial plant product comprises oil, meal, protein isolate orbiofumigant.
 19. A commercial plant product produced from the Brassicacarinata plant of claim 3, wherein the commercial plant productcomprises oil, meal, protein isolate or biofumigant.
 20. A commercialplant product produced from the F1 hybrid plant of claim 8, wherein thecommercial plant product comprises oil, meal, protein isolate orbiofumigant